52 results on '"Sebastian Sjöqvist"'
Search Results
2. RNA-sequencing reveals positional memory of multipotent mesenchymal stromal cells from oral and maxillofacial tissue transcriptomes
- Author
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Satoru Onizuka, Yasuharu Yamazaki, Sung-Joon Park, Takayuki Sugimoto, Yumiko Sone, Sebastian Sjöqvist, Michihiko Usui, Akira Takeda, Kenta Nakai, Keisuke Nakashima, and Takanori Iwata
- Subjects
Multipotent mesenchymal stromal cells ,RNA-sequencing ,HOX genes ,Maxillofacial bone ,Iliac bone ,Periodontal ligament ,Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
Abstract Background Multipotent mesenchymal stromal cells (MSCs) can be isolated from numerous tissues and are attractive candidates for therapeutic clinical applications due to their immunomodulatory and pro-regenerative capacity. Although the minimum criteria for defining MSCs have been defined, their characteristics are known to vary depending on their tissue of origin. Results We isolated and characterized human MSCs from three different bones (ilium (I-MSCs), maxilla (Mx-MSCs) and mandible (Md-MSCs)) and proceeded with next generation RNA-sequencing. Furthermore, to investigate the gene expression profiles among other cell types, we obtained RNA-seq data of human embryonic stem cells (ESCs) and several types of MSCs (periodontal ligament-derived MSCs, bone marrow-derived MSCs, and ESCs-derived MSCs) from the Sequence Reads Archive and analyzed the transcriptome profile. We found that MSCs derived from tissues of the maxillofacial region, such as the jaw bone and periodontal ligament, were HOX-negative, while those derived from other tissues were HOX-positive. We also identified that MSX1, LHX8, and BARX1, an essential regulator of craniofacial development, were strongly expressed in maxillofacial tissue-derived MSCs. Although MSCs may be divided into two distinct groups, the cells originated from over the neck or not, on the basis of differences in gene expression profile, the expression patterns of all CD antigen genes were similar among different type of MSCs, except for ESCs. Conclusions Our findings suggest that MSCs from different anatomical locations, despite meeting general characterization criteria, have remarkable differences in gene expression and positional memory. Although stromal cells from different anatomical sources are generally categorized as MSCs, their differentiation potential and biological functions vary. We suggested that MSCs may retain an original tissue memory about the developmental process, including gene expression profiles. This could have an important impact when choosing an appropriate cell source for regenerative therapy using MSCs.
- Published
- 2020
- Full Text
- View/download PDF
3. Exosomes derived from clinical-grade oral mucosal epithelial cell sheets promote wound healing
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Sebastian Sjöqvist, Taichi Ishikawa, Daisuke Shimura, Yoshiyuki Kasai, Aya Imafuku, Sophia Bou-Ghannam, Takanori Iwata, and Nobuo Kanai
- Subjects
extracellular vesicles ,exosomes ,wound healing ,oral keratinocytes ,clinical samples ,therapy ,regenerative medicine ,Cytology ,QH573-671 - Abstract
The oral mucosa exhibits unique regenerative properties, sometimes referred to as foetal-like wound healing. Researchers from our institute have used sheets of oral mucosa epithelial cells (OMECs) for regenerative medicine applications including cornea replacement and oesophageal epithelial regeneration for stricture prevention. Here, we have isolated exosomes from clinical-grade production of OMEC sheets from healthy human donors (n = 8), aiming to evaluate the clinical potential of the exosomes to stimulate epithelial regeneration and to improve understanding of the mode-of-action of the cells. Exosomes were isolated from conditioned (cExo) and non-conditioned (ncExo) media. Characterization was performed using Western blot for common exosomal-markers: CD9 and flotillin were positive while annexin V, EpCam and contaminating marker GRP94 were negative. Nanoparticle tracking analysis revealed a diameter of ~120 nm and transmission electron microscopy showed a corresponding size and spherical appearance. Human skin fibroblasts exposed to exosomes showed dose-dependent reduction of proliferation and a considerable increase of growth factor gene expression (HGF, VEGFA, FGF2 and CTGF). The results were similar for both groups, but with a trend towards a larger effect from cExo. To study adhesion, fluorescently labelled exosomes were topically applied to pig oesophageal wound-beds ex vivo and subsequently washed. Positive signal could be detected after as little as 1 min of adhesion, but increased adhesion time produced a stronger signal. Next, labelled exosomes were added to full-thickness skin wounds in rats and signal was detected up to 5 days after application. cExo significantly reduced the wound size at days 6 and 17. In conclusion, exosomes from OMEC sheets showed pro-regenerative effects on skin wound healing. This is the first time that the healing capacity of the oral mucosa is studied from an exosome perspective. These findings might lead to a combinational therapy of cell sheets and exosomes for future patients with early oesophageal cancer.
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- 2019
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4. Correction: Publisher Correction: Experimental orthotopic transplantation of a tissue-engineered oesophagus in rats
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Sebastian Sjöqvist, Philipp Jungebluth, Mei Ling Lim, Johannes C. Haag, Ylva Gustafsson, Greg Lemon, Silvia Baiguera, Miguel Angel Burguillos, Costantino Del Gaudio, Antonio Beltrán Rodríguez, Alexander Sotnichenko, Karolina Kublickiene, Henrik Ullman, Heike Kielstein, Peter Damberg, Alessandra Bianco, Rainer Heuchel, Ying Zhao, Domenico Ribatti, Cristián Ibarra, Bertrand Joseph, Doris A. Taylor, and Paolo Macchiarini
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Science - Abstract
Nature Communications 5: Article number: 3562 (2014); Published online: 15 April 2014; Updated: 10 April 2018 The original HTML version of this Article had an incorrect article number of 4562; it should have been 3562. This has now been corrected in the HTML; the PDF version of the Article was correct from the time of publication.
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- 2018
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5. Retraction Note: Experimental orthotopic transplantation of a tissue-engineered oesophagus in rats
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Sebastian Sjöqvist, Philipp Jungebluth, Mei Ling Lim, Johannes C. Haag, Ylva Gustafsson, Greg Lemon, Silvia Baiguera, Miguel Angel Burguillos, Costantino Del Gaudio, Antonio Beltrán Rodríguez, Alexander Sotnichenko, Karolina Kublickiene, Henrik Ullman, Heike Kielstein, Peter Damberg, Alessandra Bianco, Rainer Heuchel, Ying Zhao, Domenico Ribatti, Cristián Ibarra, Bertrand Joseph, Doris A. Taylor, and Paolo Macchiarini
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Science - Abstract
Nature Communications 5: Article number: 3562 (2014); Published 15 April 2014; Updated 21 March 2017 This Article is retracted by the authors. Nature Communications previously issued an Editorial Expression of Concern (http://www.nature.com/articles/ncomms13310) related to this Article, following the publication of a report commissioned by The Karolinska Institute and prepared by the Expert Group for Misconduct in Research at the Swedish Central Ethical Review Board.
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- 2017
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6. Characterization of stem-like cells in mucoepidermoid tracheal paediatric tumor.
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Mei Ling Lim, Brandon Nick Sern Ooi, Philipp Jungebluth, Sebastian Sjöqvist, Isabell Hultman, Greg Lemon, Ylva Gustafsson, Jurate Asmundsson, Silvia Baiguera, Iyadh Douagi, Irina Gilevich, Alina Popova, Johannes Cornelius Haag, Antonio Beltrán Rodríguez, Jianri Lim, Agne Liedén, Magnus Nordenskjöld, Evren Alici, Duncan Baker, Christian Unger, Tom Luedde, Ivan Vassiliev, Jose Inzunza, Lars Ahrlund-Richter, and Paolo Macchiarini
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Medicine ,Science - Abstract
Stem cells contribute to regeneration of tissues and organs. Cells with stem cell-like properties have been identified in tumors from a variety of origins, but to our knowledge there are yet no reports on tumor-related stem cells in the human upper respiratory tract. In the present study, we show that a tracheal mucoepidermoid tumor biopsy obtained from a 6 year-old patient contained a subpopulation of cells with morphology, clonogenicity and surface markers that overlapped with bone marrow mesenchymal stromal cells (BM-MSCs). These cells, designated as MEi (mesenchymal stem cell-like mucoepidermoid tumor) cells, could be differentiated towards mesenchymal lineages both with and without induction, and formed spheroids in vitro. The MEi cells shared several multipotent characteristics with BM-MSCs. However, they displayed differences to BM-MSCs in growth kinectics and gene expression profiles relating to cancer pathways and tube development. Despite this, the MEi cells did not possess in vivo tumor-initiating capacity, as proven by the absence of growth in situ after localized injection in immunocompromised mice. Our results provide an initial characterization of benign tracheal cancer-derived niche cells. We believe that this report could be of importance to further understand tracheal cancer initiation and progression as well as therapeutic development.
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- 2014
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7. Editorial Expression of Concern: Experimental orthotopic transplantation of a tissue-engineered oesophagus in rats
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Sebastian Sjöqvist, Philipp Jungebluth, Mei Ling Lim, Johannes C. Haag, Ylva Gustafsson, Greg Lemon, Silvia Baiguera, Miguel Angel Burguillos, Costantino Del Gaudio, Antonio Beltran Rodriguez, Alexander Sotnichenko, Karolina Kublickiene, Henrik Ullman, Heike Kielstein, Peter Damberg, Alessandra Bianco, Rainer Heuchel, Ying Zhao, Domenico Ribatti, Cristián Ibarra, Bertrand Joseph, Doris A. Taylor, and Paolo Macchiarini
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Science - Published
- 2016
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8. Regenerative medicine for esophageal reconstruction after cancer treatment
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Masayuki Yamato, Takeshi Ohki, Nobuo Kanai, Makoto Kondo, Susumu Eguchi, Sebastian Sjöqvist, Pontus Blomberg, and Matthius Löhr
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Medicine (General) ,R5-920 ,Cytology ,QH573-671 - Published
- 2015
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9. A pilot study using proximity extension assay of cerebrospinal fluid and its extracellular vesicles identifies novel amyotrophic lateral sclerosis biomarker candidates
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Sebastian Sjöqvist and Kentaro Otake
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Extracellular Matrix Proteins ,Extracellular Vesicles ,Amyotrophic Lateral Sclerosis ,Biophysics ,Humans ,Pilot Projects ,Cell Biology ,Molecular Biology ,Biochemistry ,Biomarkers - Abstract
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder which is characterized by progressive degeneration of the motor system. Typically, the disease starts with focal weakness which spreads to involve most muscles and leads to death from respiratory failure within five years of diagnosis. Due to the heterogenic nature of the disease, diagnostics is complex, and it generally takes twelve months from symptom-onset to diagnosis. The discovery of novel biomarkers could lead to accelerated diagnosis, earlier start of treatment, improved patient-segmentation, and treatment follow-up as well as an increased insight into the pathology. Here, we analyzed cerebrospinal fluid (CSF) and CSF-derived extracellular vesicles (CSF-EVs) from ALS-patients and matched controls (n = 9 each) using the ultra-sensitive proximity extension assay (PEA), cardiovascular III-panel. On average, 84 and 61 proteins could be detected in CSF and CSF-EVs respectively. In CSF, three proteins were significantly upregulated in ALS-patients (Junctional Adhesion Molecule A Protein, Tumor necrosis factor receptor 2 and Chitinase 1) while myoglobin was down-regulated. In CSF-EVs, no significantly differentially expressed proteins were identified, but there was a trend for downregulation of Perlecan. To our knowledge, only CHIT1 has been previously described as a CSF-based biomarker candidate for ALS. By combining the four differentially expressed markers in CSF and support vector machine algorithm, all ALS patients and 8 of 9 controls were correctly classified. In conclusion, we here demonstrate the feasibility of using PEA of CSF and CSF-EVs for biomarker discovery and propose three de novo biomarker candidates for ALS, however, further studies are necessary to demonstrate clinical usability.
- Published
- 2022
10. Analysis of Cerebrospinal Fluid Extracellular Vesicles by Proximity Extension Assay: A Comparative Study of Four Isolation Kits
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Kentaro Otake, Yoshihiko Hirozane, and Sebastian Sjöqvist
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Proteomics ,Coefficient of variation ,Nanoparticle tracking analysis ,Chemical Fractionation ,Article ,Catalysis ,cerebrospinal fluid ,lcsh:Chemistry ,Inorganic Chemistry ,Cerebrospinal fluid ,Central Nervous System Diseases ,Humans ,Physical and Theoretical Chemistry ,Biomarker discovery ,lcsh:QH301-705.5 ,Molecular Biology ,Spectroscopy ,CD63 ,Chemistry ,Organic Chemistry ,Albumin ,Cerebrospinal Fluid Proteins ,General Medicine ,Blood proteins ,Molecular biology ,Computer Science Applications ,lcsh:Biology (General) ,lcsh:QD1-999 ,proximity extension assay ,Reagent Kits, Diagnostic ,extracellular vesicles ,Biomarkers ,CD81 - Abstract
There is a lack of reliable biomarkers for disorders of the central nervous system (CNS), and diagnostics still heavily rely on symptoms that are both subjective and difficult to quantify. The cerebrospinal fluid (CSF) is a promising source of biomarkers due to its close connection to the CNS. Extracellular vesicles are actively secreted by cells, and proteomic analysis of CSF extracellular vesicles (EVs) and their molecular composition likely reflects changes in the CNS to a higher extent compared with total CSF, especially in the case of neuroinflammation, which could increase blood&ndash, brain barrier permeability and cause an influx of plasma proteins into the CSF. We used proximity extension assay for proteomic analysis due to its high sensitivity. We believe that this methodology could be useful for de novo biomarker discovery for several CNS diseases. We compared four commercially available kits for EV isolation: MagCapture and ExoIntact (based on magnetic beads), EVSecond L70 (size-exclusion chromatography), and exoEasy (membrane affinity). The isolated EVs were characterized by nanoparticle tracking analysis, ELISA (CD63, CD81 and albumin), and proximity extension assay (PEA) using two different panels, each consisting of 92 markers. The exoEasy samples did not pass the built-in quality controls and were excluded from downstream analysis. The number of detectable proteins in the ExoIntact samples was considerably higher (~150% for the cardiovascular III panel and ~320% for the cell regulation panel) compared with other groups. ExoIntact also showed the highest intersample correlation with an average Pearson&rsquo, s correlation coefficient of 0.991 compared with 0.985 and 0.927 for MagCapture and EVSecond, respectively. The median coefficient of variation was 5%, 8%, and 22% for ExoIntact, MagCapture, and EVSecond, respectively. Comparing total CSF and ExoIntact samples revealed 70 differentially expressed proteins in the cardiovascular III panel and 17 in the cell regulation panel. To our knowledge, this is the first time that CSF EVs were analyzed by PEA. In conclusion, analysis of CSF EVs by PEA is feasible, and different isolation kits give distinct results, with ExoIntact showing the highest number of identified proteins with the lowest variability.
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- 2020
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11. Correction to: RNA-sequencing reveals positional memory of multipotent mesenchymal stromal cells from oral and maxillofacial tissue transcriptomes
- Author
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Michihiko Usui, Keisuke Nakashima, Takanori Iwata, Sebastian Sjöqvist, Kenta Nakai, Sung-Joon Park, Yumiko Sone, Yasuharu Yamazaki, Akira Takeda, Takayuki Sugimoto, and Satoru Onizuka
- Subjects
Iliac bone ,lcsh:QH426-470 ,Maxillofacial bone ,lcsh:Biotechnology ,RNA-sequencing ,RNA ,Multipotent Mesenchymal Stromal Cells ,Computational biology ,Biology ,Proteomics ,HOX genes ,Transcriptome ,lcsh:Genetics ,lcsh:TP248.13-248.65 ,Genetics ,DNA microarray ,Multipotent mesenchymal stromal cells ,Biotechnology ,Research Article ,Periodontal ligament - Abstract
Background Multipotent mesenchymal stromal cells (MSCs) can be isolated from numerous tissues and are attractive candidates for therapeutic clinical applications due to their immunomodulatory and pro-regenerative capacity. Although the minimum criteria for defining MSCs have been defined, their characteristics are known to vary depending on their tissue of origin. Results We isolated and characterized human MSCs from three different bones (ilium (I-MSCs), maxilla (Mx-MSCs) and mandible (Md-MSCs)) and proceeded with next generation RNA-sequencing. Furthermore, to investigate the gene expression profiles among other cell types, we obtained RNA-seq data of human embryonic stem cells (ESCs) and several types of MSCs (periodontal ligament-derived MSCs, bone marrow-derived MSCs, and ESCs-derived MSCs) from the Sequence Reads Archive and analyzed the transcriptome profile. We found that MSCs derived from tissues of the maxillofacial region, such as the jaw bone and periodontal ligament, were HOX-negative, while those derived from other tissues were HOX-positive. We also identified that MSX1, LHX8, and BARX1, an essential regulator of craniofacial development, were strongly expressed in maxillofacial tissue-derived MSCs. Although MSCs may be divided into two distinct groups, the cells originated from over the neck or not, on the basis of differences in gene expression profile, the expression patterns of all CD antigen genes were similar among different type of MSCs, except for ESCs. Conclusions Our findings suggest that MSCs from different anatomical locations, despite meeting general characterization criteria, have remarkable differences in gene expression and positional memory. Although stromal cells from different anatomical sources are generally categorized as MSCs, their differentiation potential and biological functions vary. We suggested that MSCs may retain an original tissue memory about the developmental process, including gene expression profiles. This could have an important impact when choosing an appropriate cell source for regenerative therapy using MSCs.
- Published
- 2020
12. RNA-sequencing reveals positional memory of multipotent mesenchymal stromal cells from oral and maxillofacial tissue transcriptomes
- Author
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Yumiko Sone, Michihiko Usui, Sung-Joon Park, Sebastian Sjöqvist, Takanori Iwata, Keisuke Nakashima, Yasuharu Yamazaki, Kenta Nakai, Akira Takeda, Takayuki Sugimoto, and Satoru Onizuka
- Subjects
Iliac bone ,Cell type ,Stromal cell ,Maxillofacial bone ,lcsh:QH426-470 ,lcsh:Biotechnology ,RNA-sequencing ,Mandible ,Biology ,Regenerative medicine ,Ilium ,Transcriptome ,03 medical and health sciences ,lcsh:TP248.13-248.65 ,Exome Sequencing ,Gene expression ,Maxilla ,Genetics ,Humans ,Periodontal fiber ,Gene Regulatory Networks ,Multipotent mesenchymal stromal cells ,Cells, Cultured ,Cell Proliferation ,030304 developmental biology ,Homeodomain Proteins ,0303 health sciences ,Sequence Analysis, RNA ,Gene Expression Profiling ,030302 biochemistry & molecular biology ,Mesenchymal stem cell ,High-Throughput Nucleotide Sequencing ,Correction ,Cell Differentiation ,Mesenchymal Stem Cells ,HOX genes ,Embryonic stem cell ,Cell biology ,lcsh:Genetics ,Gene Expression Regulation ,Organ Specificity ,Periodontal ligament ,Biotechnology - Abstract
Background Multipotent mesenchymal stromal cells (MSCs) can be isolated from numerous tissues and are attractive candidates for therapeutic clinical applications due to their immunomodulatory and pro-regenerative capacity. Although the minimum criteria for defining MSCs have been defined, their characteristics are known to vary depending on their tissue of origin. Results We isolated and characterized human MSCs from three different bones (ilium (I-MSCs), maxilla (Mx-MSCs) and mandible (Md-MSCs)) and proceeded with next generation RNA-sequencing. Furthermore, to investigate the gene expression profiles among other cell types, we obtained RNA-seq data of human embryonic stem cells (ESCs) and several types of MSCs (periodontal ligament-derived MSCs, bone marrow-derived MSCs, and ESCs-derived MSCs) from the Sequence Reads Archive and analyzed the transcriptome profile. We found that MSCs derived from tissues of the maxillofacial region, such as the jaw bone and periodontal ligament, were HOX-negative, while those derived from other tissues were HOX-positive. We also identified that MSX1, LHX8, and BARX1, an essential regulator of craniofacial development, were strongly expressed in maxillofacial tissue-derived MSCs. Although MSCs may be divided into two distinct groups, the cells originated from over the neck or not, on the basis of differences in gene expression profile, the expression patterns of all CD antigen genes were similar among different type of MSCs, except for ESCs. Conclusions Our findings suggest that MSCs from different anatomical locations, despite meeting general characterization criteria, have remarkable differences in gene expression and positional memory. Although stromal cells from different anatomical sources are generally categorized as MSCs, their differentiation potential and biological functions vary. We suggested that MSCs may retain an original tissue memory about the developmental process, including gene expression profiles. This could have an important impact when choosing an appropriate cell source for regenerative therapy using MSCs.
- Published
- 2020
13. Extracellular Vesicle Therapeutics in Regenerative Medicine
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Aya Imafuku and Sebastian Sjöqvist
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Chemistry ,Vesicle ,Extracellular vesicle ,medicine.disease ,Regenerative medicine ,Cell biology ,Cell therapy ,03 medical and health sciences ,0302 clinical medicine ,Graft-versus-host disease ,medicine ,Nucleic acid ,030212 general & internal medicine ,Induced pluripotent stem cell ,Intracellular - Abstract
Extracellular vesicles (EVs) are nano-sized, cell-released vesicles which contain lipids, proteins, and nucleic acids derived from the parental cells. EVs play an important role in intercellular communication and influence both physiological and pathological conditions. They are increasingly explored as potential therapeutic agents since they can cross biological barriers, their cargo is protected from degradation and they are involved in the transfer of bioactive components. EVs can promote tissue regeneration and might be alternatives to cell therapy. They can be used both in their native form, and as delivery vehicles for therapeutic agents. However, there are many hurdles to overcome for broad clinical application of EVs as therapeutics. Here, we review recent conditions regarding EVs therapeutics in regenerative medicine.
- Published
- 2020
14. Transplantation of tissue‐engineered cell sheets for stricture prevention after endoscopic submucosal dissection of the oesophagus
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Eduard Jonas, Magnus Nilsson, Mei Ling Lim, Pontus Blomberg, Peter Elbe, Teruo Okano, Makoto Kondo, Stephan L. Haas, J-Matthias Löhr, Jenny Enger, Takeshi Ohki, Johan Permert, Masayuki Yamato, Nobuo Kanai, Katrin Markland, Ryo Takagi, Sebastian Sjöqvist, Masakazu Yamamoto, and Ammar Mohkles-Barakat
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medicine.medical_specialty ,Tissue engineered ,business.industry ,Gastroenterology ,Treatment options ,Original Articles ,Endoscopic submucosal dissection ,Dissection (medical) ,medicine.disease ,digestive system diseases ,Surgery ,Transplantation ,03 medical and health sciences ,0302 clinical medicine ,Oncology ,030220 oncology & carcinogenesis ,Barrett's oesophagus ,medicine ,030211 gastroenterology & hepatology ,business ,Cell sheet - Abstract
Endoscopic mucosal dissection (ESD) is a treatment option for oesophagus tumours localized to the mucosa enabling en bloc removal of large lesions. The resulting larger mucosal defects have resulted in an increase in the occurrence of post-treatment strictures. Transplantation of autologous cell sheets, cultured from oral mucosa, has been shown to prevent post-ESD strictures. The aim of the study was to assess the efficacy and safety of cell sheet transplantation after oesophageal ESD in a Western patient population where reflux-associated pre-malignant and malignant conditions predominate.Patients with Barrett's oesophagus associated high-grade dysplasia or early adenocarcinoma where ESD entailed a resection3 cm in length and ≥75% of the circumference were eligible for treatment under hospital exemption. Cell sheets were cultured from buccal mucosa according to Good Manufacturing Practice and were endoscopically applied to the post-ESD defect directly after resection. Patients were followed with weekly endoscopy examinations, including confocal laser microscopy, for a total of four weeks.Five patients were treated. ESD was extensive with resections being circumferential in three patients and 9-10 cm in length in two. The number of transplanted cell sheets ranged from two to six. Three patients developed strictures requiring two to five dilatation sessions.Cell sheet transplantation shows to be safe and feasible in a Western population. Results suggest that transplantation has a protective effect on the mucosal defect after ESD, decreasing both the risk for and extent of stricture formation.
- Published
- 2016
15. The Use of Mathematical Modelling for Improving the Tissue Engineering of Organs and Stem Cell Therapy
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Annika Stuewer, Philipp Jungebluth, Alexandra B. Firsova, Paolo Macchiarini, Johannes C. Haag, Risul Amin, E. A. Gubareva, Sebastian Sjöqvist, Mei Ling Lim, Greg Lemon, Neus Feliu, and Ylva Gustafsson
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,medicine.medical_treatment ,Science and engineering ,0206 medical engineering ,Medicine (miscellaneous) ,02 engineering and technology ,Biology ,Regenerative Medicine ,Models, Biological ,Regenerative medicine ,03 medical and health sciences ,Tissue scaffolds ,Tissue engineering ,medicine ,Animals ,Humans ,Tissue Engineering ,Tissue Scaffolds ,Management science ,General Medicine ,Stem-cell therapy ,030104 developmental biology ,Organ Specificity ,020602 bioinformatics ,Stem Cell Transplantation - Abstract
Regenerative medicine is a multidisciplinary field where continued progress relies on the incorporation of a diverse set of technologies from a wide range of disciplines within medicine, science and engineering. This review describes how one such technique, mathematical modelling, can be utilised to improve the tissue engineering of organs and stem cell therapy. Several case studies, taken from research carried out by our group, ACTREM, demonstrate the utility of mechanistic mathematical models to help aid the design and optimisation of protocols in regenerative medicine.
- Published
- 2016
16. Isolation and Characterization of Extracellular Vesicles from Keratinocyte Cultures
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Sebastian, Sjöqvist, Aya, Imafuku, Dhanu, Gupta, and Samir, El Andaloussi
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Keratinocytes ,Melanins ,Extracellular Vesicles ,Microscopy, Electron, Transmission ,Culture Media, Conditioned ,Blotting, Western ,Humans ,Particle Size ,Cells, Cultured ,Cell Line - Abstract
Extracellular vesicles (EVs), including exosomes, are nano-sized membrane-bound particles which are released by cells. They have been found in all examined body fluids and can be isolated from conditioned cell culture media. These vesicles have gained increasing attention due to their importance in cellular cross talk, in both health and disease. For example, keratinocyte-derived EVs have been described to modulate melanin production in epidermis. Similar EVs were also shown to have an important role in skin immunology, by stimulating dendritic cells. In this chapter, we will describe how to isolate EVs from keratinocyte cultures and how to perform characterization by Western blot, nanoparticle tracking analysis, and transmission electron microscopy.
- Published
- 2019
17. Oral keratinocyte-derived exosomes regulate proliferation of fibroblasts and epithelial cells
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Sebastian Sjöqvist, Nadiah Ali, Daisuke Shimura, Yoshiyuki Kasai, Nobuo Kanai, Takanori Iwata, and Taichi Ishikawa
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0301 basic medicine ,Keratinocytes ,Cell ,Biophysics ,Exosomes ,Biochemistry ,Cell Line ,Dermal fibroblast ,03 medical and health sciences ,0302 clinical medicine ,Western blot ,Annexin ,medicine ,Humans ,Oral mucosa ,Particle Size ,Molecular Biology ,Cell Proliferation ,medicine.diagnostic_test ,Cell growth ,Chemistry ,Epithelial Cells ,Cell Biology ,Fibroblasts ,Microvesicles ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Keratinocyte - Abstract
Extracellular vesicles (EVs), including exosomes, are small membrane-bound particles released by cells. From a therapeutic point of view, EVs can often convey similar biological function as their parent cell. Grafts originating from oral mucosa have frequently been used in regenerative medicine, and we have previously described the use of oral cell sheets to prevent stricture formation of the esophagus. Further, we recently found that exosomes derived from these cell sheets have pro-regenerative effect on skin wound healing. Here, we have isolated exosomes from conditioned media from oral keratinocyte (“OKEx”) and dermal fibroblast (“FEx”) cultures. The exosomes were probed for classical EV-markers by western blot (CD9, annexin V and Flotillin-1), FEx were positive for all markers while OKEx were positive only for CD9. Tunable resistive pulse sensing indicated a mean size of around 110 nm and transmission electron microscopy showed a spherical morphology, for both groups. After fluorescent labelling, we studied the uptake of exosomes co-cultured with fibroblasts or keratinocytes. Signal from OKEx could be detected after 90 min, and signal could be detected in all groups after 16 h. Finally we studied the exosomes’ modulation of cell proliferation. Both groups suppressed proliferation of healthy keratinocyte and fibroblasts, at some doses to similar levels as dexamethasone (a drug commonly used to prevent stricture formation). In contrast, the exosomes also suppressed the proliferation of the carcinoma cell line TR146, while dexamethasone had no effect. In conclusion, we believe that exosome-signaling might be one of the mode-of-actions of cell sheet-therapy for stricture prevention.
- Published
- 2019
18. Retraction notice to:'Verification of cell viability in bioengineered tissues and organs before clinical transplantation ' [BIOMATERIALS (2013) 4057-4067]
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Oscar E. Simonson, Sebastian Sjöqvist, Greg Lemon, Ylva Gustafsson, Karl H. Grinnemo, Costantino Del Gaudio, Mei Ling Lim, Staffan Strömblad, Johannes C. Haag, Paolo Macchiarini, I. V. Gilevich, Philipp Jungebluth, Matthias Corbascio, Fatemeh Ajalloueian, and Silvia Baiguera
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Biomaterials ,Transplantation ,Pathology ,medicine.medical_specialty ,Notice ,Mechanics of Materials ,business.industry ,Biophysics ,Ceramics and Composites ,Medicine ,Bioengineering ,Viability assay ,business - Published
- 2019
19. Correction to: Isolation and Characterization of Extracellular Vesicles from Keratinocyte Cultures
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Aya Imafuku, Sebastian Sjöqvist, Dhanu Gupta, and Samir El Andaloussi
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medicine.anatomical_structure ,medicine ,Biology ,Isolation (microbiology) ,Keratinocyte ,Molecular biology ,Extracellular vesicles - Abstract
The original version of this chapter was inadvertently published with incorrect spelling of surname of the authors. The names should read Sebastian Sjoqvist, Aya Imafuku, Danu Gupta, and Samir EL Andaloussi, and not Sebastian Sjoqvist, Aya Imafuku, Dhanu Ghupta, and Samir E. L. Andaloussi.
- Published
- 2019
20. Exosomes derived from clinical-grade oral mucosal epithelial cell sheets promote wound healing
- Author
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Sophia Bou-Ghannam, Nobuo Kanai, Daisuke Shimura, Taichi Ishikawa, Yoshiyuki Kasai, Aya Imafuku, Sebastian Sjöqvist, and Takanori Iwata
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,Histology ,regenerative medicine ,wound healing ,exosomes ,Regenerative medicine ,Extracellular vesicles ,03 medical and health sciences ,0302 clinical medicine ,oral keratinocytes ,medicine ,Oral mucosa ,lcsh:QH573-671 ,therapy ,business.industry ,lcsh:Cytology ,clinical samples ,Oral mucosal epithelial cell ,Clinical grade ,Cell Biology ,Microvesicles ,stomatognathic diseases ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,business ,Wound healing ,Research Article - Abstract
The oral mucosa exhibits unique regenerative properties, sometimes referred to as foetal-like wound healing. Researchers from our institute have used sheets of oral mucosa epithelial cells (OMECs) for regenerative medicine applications including cornea replacement and oesophageal epithelial regeneration for stricture prevention. Here, we have isolated exosomes from clinical-grade production of OMEC sheets from healthy human donors (n = 8), aiming to evaluate the clinical potential of the exosomes to stimulate epithelial regeneration and to improve understanding of the mode-of-action of the cells. Exosomes were isolated from conditioned (cExo) and non-conditioned (ncExo) media. Characterization was performed using Western blot for common exosomal-markers: CD9 and flotillin were positive while annexin V, EpCam and contaminating marker GRP94 were negative. Nanoparticle tracking analysis revealed a diameter of ~120 nm and transmission electron microscopy showed a corresponding size and spherical appearance. Human skin fibroblasts exposed to exosomes showed dose-dependent reduction of proliferation and a considerable increase of growth factor gene expression (HGF, VEGFA, FGF2 and CTGF). The results were similar for both groups, but with a trend towards a larger effect from cExo. To study adhesion, fluorescently labelled exosomes were topically applied to pig oesophageal wound-beds ex vivo and subsequently washed. Positive signal could be detected after as little as 1 min of adhesion, but increased adhesion time produced a stronger signal. Next, labelled exosomes were added to full-thickness skin wounds in rats and signal was detected up to 5 days after application. cExo significantly reduced the wound size at days 6 and 17. In conclusion, exosomes from OMEC sheets showed pro-regenerative effects on skin wound healing. This is the first time that the healing capacity of the oral mucosa is studied from an exosome perspective. These findings might lead to a combinational therapy of cell sheets and exosomes for future patients with early oesophageal cancer.
- Published
- 2019
21. RNA-Sequencing Reveals Unique Features of Multipotent Mesenchymal Stromal Cells from Oral and Maxillofacial Tissue Transcriptomes
- Author
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Kenta Nakai, Michihiko Usui, Sung-Joon Park, Takayuki Sugimoto, Satoru Onizuka, Yasuharu Yamazaki, Takanori Iwata, Sebastian Sjöqvist, Keisuke Nakashima, Yumiko Sone, and Akira Takeda
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Transcriptome ,Cell type ,Stromal cell ,Mesenchymal stem cell ,Biology ,Hox gene ,Medical research ,Bioinformatics ,Gene ,Regenerative medicine - Abstract
Background: Multipotent mesenchymal stromal cells (MSCs) can be isolated from numerous tissues and are attractive candidates for therapeutic clinical applications due to their immunomodulatory and pro-regenerative capacity. Although the minimum criteria for defining MSCs have been defined, their characteristics are known to vary depending on their tissue of origin. Methods: We isolated and characterized human MSCs from three different bones (ilium (I-MSCs), maxilla (Mx-MSCs) and mandibular (Md-MSCs)) and proceeded with next generation RNA-sequencing. Furthermore, to investigate the gene expression profiles among other cell types, we obtained RNA-seq data of numerous types of MSCs from the Sequence Reads Archive and analyze the transcriptome profile. Findings: We analyzed RNA-sequencing data of several MSC types and found that those from tissues of the maxillofacial region, such as the jaw bone and periodontal ligament, were HOX-negative, while those from other tissues were HOX-positive. We also identified the genes strongly expressed in maxillofacial tissue-derived MSCs were involved in the craniofacial development. Interpretation: Our findings suggest that MSCs from different anatomical locations, despite meeting general characterization criteria, have remarkable differences in gene expression and positional memory. Although stromal cells from different anatomical sources are generally categorized as MSCs, their differentiation potential and biological functions vary. We suggested that MSCs may retain a memory of the developmental process, including gene expression profiles. This could have important impact when choosing appropriate cell source for regenerative therapy using MSCs. Funding: This study was supported by the Japan Agency for Medical Research and Development and the Japan Society for the Promotion of Science. Declaration of Interest: No authors have potential conflicts of interest, including financial interests or relationships or affiliations, relevant to the subject of this manuscript. Ethical Approval: This study was approved by the Ethics Committee at Tokyo Women’s Medical University and Kitasato University that examines the use of tissues of human cell origin, and was conducted after the acquisition of written informed consent from patients or their families.
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- 2019
22. Isolation and Characterization of Extracellular Vesicles from Keratinocyte Cultures
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Samir El Andaloussi, Dhanu Gupta, Sebastian Sjöqvist, and Aya Imafuku
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0301 basic medicine ,medicine.diagnostic_test ,Epidermis (botany) ,Chemistry ,Vesicle ,Nanoparticle tracking analysis ,Isolation (microbiology) ,Microvesicles ,Cell biology ,Melanin ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,Western blot ,030220 oncology & carcinogenesis ,medicine ,Keratinocyte - Abstract
Extracellular vesicles (EVs), including exosomes, are nano-sized membrane-bound particles which are released by cells. They have been found in all examined body fluids and can be isolated from conditioned cell culture media. These vesicles have gained increasing attention due to their importance in cellular cross talk, in both health and disease. For example, keratinocyte-derived EVs have been described to modulate melanin production in epidermis. Similar EVs were also shown to have an important role in skin immunology, by stimulating dendritic cells. In this chapter, we will describe how to isolate EVs from keratinocyte cultures and how to perform characterization by Western blot, nanoparticle tracking analysis, and transmission electron microscopy.
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- 2019
23. Publisher Correction: Experimental orthotopic transplantation of a tissue-engineered oesophagus in rats
- Author
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Johannes C. Haag, Greg Lemon, Philipp Jungebluth, Silvia Baiguera, Cristian Ibarra, Paolo Macchiarini, Rainer Heuchel, Karolina Kublickiene, Doris A. Taylor, Peter Damberg, Bertrand Joseph, Alessandra Bianco, Domenico Ribatti, Antonio B. Rodriguez, Ylva Gustafsson, Miguel Angel Burguillos, Sebastian Sjöqvist, Mei Ling Lim, Costantino Del Gaudio, Alexander Sotnichenko, Ying Zhao, Heike Kielstein, and Henrik Ullman
- Subjects
medicine.medical_specialty ,Science ,Myocytes, Smooth Muscle ,MEDLINE ,General Physics and Astronomy ,General Biochemistry, Genetics and Molecular Biology ,Orthotopic transplantation ,Esophagus ,Medicine ,Animals ,Regeneration ,Multidisciplinary ,Tissue engineered ,Tissue Engineering ,Tissue Scaffolds ,business.industry ,Published Erratum ,General surgery ,Correction ,Cell Differentiation ,Mesenchymal Stem Cells ,General Chemistry ,Rats ,business ,Immunocompetence - Abstract
A tissue-engineered oesophageal scaffold could be very useful for the treatment of pediatric and adult patients with benign or malignant diseases such as carcinomas, trauma or congenital malformations. Here we decellularize rat oesophagi inside a perfusion bioreactor to create biocompatible biological rat scaffolds that mimic native architecture, resist mechanical stress and induce angiogenesis. Seeded allogeneic mesenchymal stromal cells spontaneously differentiate (proven by gene-, protein and functional evaluations) into epithelial- and muscle-like cells. The reseeded scaffolds are used to orthotopically replace the entire cervical oesophagus in immunocompetent rats. All animals survive the 14-day study period, with patent and functional grafts, and gain significantly more weight than sham-operated animals. Explanted grafts show regeneration of all the major cell and tissue components of the oesophagus including functional epithelium, muscle fibres, nerves and vasculature. We consider the presented tissue-engineered oesophageal scaffolds a significant step towards the clinical application of bioengineered oesophagi.
- Published
- 2018
24. Retraction Note: Experimental orthotopic transplantation of a tissue-engineered oesophagus in rats
- Author
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Silvia Baiguera, Johannes C. Haag, Henrik Ullman, Bertrand Joseph, Paolo Macchiarini, Alessandra Bianco, Doris A. Taylor, Karolina Kublickiene, Costantino Del Gaudio, Mei Ling Lim, Domenico Ribatti, Ying Zhao, Ylva Gustafsson, Philipp Jungebluth, Heike Kielstein, Cristian Ibarra, Antonio Beltrán Rodríguez, Alexander Sotnichenko, Rainer Heuchel, Peter Damberg, Greg Lemon, Miguel Angel Burguillos, and Sebastian Sjöqvist
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Multidisciplinary ,Tissue engineered ,business.industry ,Science ,General Physics and Astronomy ,General Chemistry ,General Biochemistry, Genetics and Molecular Biology ,Retraction ,03 medical and health sciences ,030104 developmental biology ,Orthotopic transplantation ,Medicine ,business - Abstract
Nature Communications 5: Article number: 3562 (2014); Published 15 April 2014; Updated 21 March 2017 This Article is retracted by the authors. Nature Communications previously issued an Editorial Expression of Concern (http://www.nature.com/articles/ncomms13310) related to this Article, following the publication of a report commissioned by The Karolinska Institute and prepared by the Expert Group for Misconduct in Research at the Swedish Central Ethical Review Board.
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- 2017
25. Diverse applications of nanomedicine
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Philipp Jungebluth, Ali Khademhosseini, Xian-En Zhang, Yuzhou Wu, Tai Hyun Park, Christian Dullin, Helmuth Möhwald, Neus Feliu, Mahmoud Soliman, Michael D. George, Nicholas A. Kotov, Buddhisha Udugama, Paul Mulvaney, Ramon A. Alvarez-Puebla, Warren C. W. Chan, Kazunori Kataoka, Sumaira Ashraf, Beatriz Pelaz, Xingyu Jiang, Yury Gogotsi, Naomi J. Halas, Yuliang Zhao, Arnold Grünweller, Laura Ballerini, Jose Oliveira, Ben Zhong Tang, Sebastian Sjöqvist, Susanna Bosi, Andre G. Skirtach, Anne M. Andrews, Teruo Okano, Daxiang Cui, Shuming Nie, Maurizio Prato, Qian Zhang, Patrick Hunziker, Alberto Escudero, Xin Zhou, Qiang Zhang, Huan Meng, Claus-Michael Lehr, Christoph Alexiou, Youqing Shen, Wolfgang J. Parak, Luis M. Liz-Marzán, Lajos P. Balogh, Ji Jian, Andre E. Nel, Molly M. Stevens, Xiaowei Ma, Paul S. Weiss, Zhao Yue, Rainer Tietze, Xiaodong Chen, Raymond E. Schaak, Zhongwei Gu, Chunying Chen, Hsing-Wen Sung, Jindřich Kopeček, Xing-Jie Liang, Alessandra Bestetti, Lily Yang, Harald F. Krug, Paolo Macchiarini, Mei Ling Lim, Vincent M. Rotello, Mónica Carril, Tanja Weil, Zhen Cheng, Pranav Kadhiresan, J. Scott VanEpps, Roland K. Hartmann, Mark C. Hersam, Xiaoyuan Chen, Itamar Willner, Mingyuan Gao, Dong Soo Lee, Amila Samarakoon, Peter Nordlander, Norbert Hampp, Víctor F. Puntes, Cornelia Brendel, Reginald M. Penner, Kam W. Leong, Jianzhong Du, Frauke Alves, Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS),Saarland 9 University, 66123 Saarbrücken, Germany., Pelaz, Beatriz, Alexiou, Christoph, Alvarez Puebla, Ramon A., Alves, Frauke, Andrews, Anne M., Ashraf, Sumaira, Balogh, Lajos P., Ballerini, Laura, Bestetti, Alessandra, Brendel, Cornelia, Bosi, Susanna, Carril, Monica, Chan, Warren C. W., Chen, Chunying, Chen, Xiaodong, Chen, Xiaoyuan, Cheng, Zhen, Cui, Daxiang, Du, Jianzhong, Dullin, Christian, Escudero, Alberto, Feliu, Neu, Gao, Mingyuan, George, Michael, Gogotsi, Yury, Grünweller, Arnold, Gu, Zhongwei, Halas, Naomi J., Hampp, Norbert, Hartmann, Roland K., Hersam, Mark C., Hunziker, Patrick, Jian, Ji, Jiang, Xingyu, Jungebluth, Philipp, Kadhiresan, Pranav, Kataoka, Kazunori, Khademhosseini, Ali, Kopeček, Jindřich, Kotov, Nicholas A., Krug, Harald F., Lee, Dong Soo, Lehr, Claus Michael, Leong, Kam W., Liang, Xing Jie, Ling Lim, Mei, Liz Marzán, Luis M., Ma, Xiaowei, Macchiarini, Paolo, Meng, Huan, Möhwald, Helmuth, Mulvaney, Paul, Nel, Andre E., Nie, Shuming, Nordlander, Peter, Okano, Teruo, Oliveira, Jose, Park, Tai Hyun, Penner, Reginald M., Prato, Maurizio, Puntes, Victor, Rotello, Vincent M., Samarakoon, Amila, Schaak, Raymond E., Shen, Youqing, Sjöqvist, Sebastian, Skirtach, Andre G., Soliman, Mahmoud G., Stevens, Molly M., Sung, Hsing Wen, Tang, Ben Zhong, Tietze, Rainer, Udugama, Buddhisha N., Vanepps, J. Scott, Weil, Tanja, Weiss, Paul S., Willner, Itamar, Wu, Yuzhou, Yang, Lily, Yue, Zhao, Zhang, Qian, Zhang, Qiang, Zhang, Xian En, Zhao, Yuliang, Zhou, Xin, Parak, Wolfgang J., German Academic Exchange Service, Chinese Academy of Sciences, National Natural Science Foundation of China, National Basic Research Program (China), European Commission, Ministerio de Economía y Competitividad (España), Generalitat de Catalunya, Swiss National Science Foundation, Julian Schwinger Foundation, Claude Leon Foundation, National Science Foundation (US), Canadian Institutes of Health Research, Natural Sciences and Engineering Research Council of Canada, Alexander von Humboldt Foundation, Lars Hierta Memorial Foundation, Eusko Jaurlaritza, Research Grants Council (Hong Kong), National Cancer Institute (US), Junta de Andalucía, Research Foundation - Flanders, and German Research Foundation
- Subjects
Technology ,Chemistry, Multidisciplinary ,neurons ,General Physics and Astronomy ,02 engineering and technology ,Settore BIO/09 - Fisiologia ,01 natural sciences ,Engineering (all) ,Drug Delivery Systems ,Imaging tools ,Neoplasms ,Medicine and Health Sciences ,Nanotechnology ,General Materials Science ,Diverse applications ,nanomaterials ,Wearable technology ,Drug Carriers ,Chemistry, Physical ,General Engineering ,021001 nanoscience & nanotechnology ,Wearable devices ,3. Good health ,Chemistry ,Nanomedicine ,Physical Sciences ,QUANTUM-DOT BARCODES ,Science & Technology - Other Topics ,Medicine ,Materials Science (all) ,0210 nano-technology ,Nano Focus ,Materials science ,Materials Science ,Physics and Astronomy (all) ,Materials Science, Multidisciplinary ,010402 general chemistry ,MESENCHYMAL STEM-CELLS ,Vaccine development ,TARGETED DRUG-DELIVERY ,LABEL-FREE DETECTION ,MESOPOROUS SILICA NANOPARTICLES ,High throughput screening ,MD Multidisciplinary ,Animals ,Humans ,SURFACE-PLASMON RESONANCE ,Nanoscience & Nanotechnology ,Particle Size ,cell physiology ,FIELD-EFFECT TRANSISTOR ,Biomedicine ,Science & Technology ,carbon nanotubes ,business.industry ,COATED GOLD NANOPARTICLES ,neurology ,IRON-OXIDE NANOPARTICLES ,Biology and Life Sciences ,Data science ,nanomedicine, neurology, nanomaterials, carbon nanotubes, cell physiology, neurons ,0104 chemical sciences ,Physics and Astronomy ,Targeted drug delivery ,Nanoscale size ,Nanoparticles ,ENHANCED RAMAN-SCATTERING ,Drug Delivery ,business - Abstract
The design and use of materials in the nanoscale size range for addressing medical and health-related issues continues to receive increasing interest. Research in nanomedicine spans a multitude of areas, including drug delivery, vaccine development, antibacterial, diagnosis and imaging tools, wearable devices, implants, high-throughput screening platforms, etc. using biological, nonbiological, biomimetic, or hybrid materials. Many of these developments are starting to be translated into viable clinical products. Here, we provide an overview of recent developments in nanomedicine and highlight the current challenges and upcoming opportunities for the field and translation to the clinic., This work was supported by the Deutscher Akademischer Austauschdienst (DAAD to Philipps Universität Marburg and Zhejiang University, Hangzhou), the Chinesisch Deutsches Zentrum für Wissenschaftsförderung (“CDZ” to Z.G. and W.J.P.), and the Chinese Academy of Science (CAS). Part of this work was supported by the National Natural Science Foundation (51390481, 81227902, 81625011), National Basic Research Program (2014CB931900) of China (to Y.S.), by the European Commission grant Futurenanoneeds (to V.P. and W.J.P.), by the Spanish Ministerio de Economia y Competitividad (CTQ2011-23167 and CTQ2014-59808R to R.A.A.P.), the Generalitat of Catalunya (2014-SGR-612 to R.A.A.P.), the Deutsche Forschungsgemeinschaft (DFG) (AL552/8-1 to R.T.), the Swiss National Science Foundation (NRP62 to P.H.), the Claude & Julianna Foundation (grant to P.H.), the National Science Foundation (NSF) grants CHE-1306928 (to R.P.) and ECS-0601345; CBET 0933384; CBET 0932823; and CBET 1036672 (to N.A.K.), Canadian Institute of Health Research (grant to W.C.W.C.), and Natural Sciences and Engineering Research Council of Canada (grant to W.C.W.C.). S.A. and B.P. acknowledge a fellowship from the Alexander von Humboldt Foundation. N.F. acknowledges the Lars Hiertas Minne Foundation. M.C. acknowledges Ikerbasque for a Research Fellow position. X.C. acknowledges the Intramural Research Program (IRP), National Institute of Biomedical Imaging and Bioengineering (NIBIB), National Institutes of Health (NIH). B.Z.T. acknowledges the Innovation and Technology Commission of Hong Kong (ITC-CNERC14SC01). The Pancreatic Cancer research of A.E.N. and H.M. was funded by the U.S. National Cancer Institute, NIH grant # U01CA198846. A.E. acknowledges Junta de Andalucía (Spain) for a Talentia Postdoc Fellowship, co-financed by the European Union's Seventh Framework Programme, grant agreement no 267226. A.G.S. acknowledges support by BOF (UGent) and FWO (Research Foundation Flanders). Part of this work was supported by the National Natural Science Foundation of China.
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- 2017
26. Tracheal tissue engineering in rats
- Author
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Philipp Jungebluth, Paolo Macchiarini, Greg Lemon, Johannes C. Haag, Ivar Dehnisch, Alessandra Bianco, Per Uhlén, Mei Ling Lim, Silvia Baiguera, Antonio Beltrán Rodríguez, Ylva Gustafsson, Costantino Del Gaudio, and Sebastian Sjöqvist
- Subjects
Decellularization ,Tissue Engineering ,Tissue Scaffolds ,Guided Tissue Regeneration ,business.industry ,Settore ING-IND/22 - Scienza e Tecnologia dei Materiali ,Cellular differentiation ,Regeneration (biology) ,Rat model ,Nanofibers ,General Biochemistry, Genetics and Molecular Biology ,Biomechanical Phenomena ,Rats ,Trachea ,Orthotopic transplantation ,Tissue engineering ,Electrospun nanofibers ,Animals ,Medicine ,Colorimetry ,Viability assay ,business ,Biomedical engineering - Abstract
Tissue-engineered tracheal transplants have been successfully performed clinically. However, before becoming a routine clinical procedure, further preclinical studies are necessary to determine the underlying mechanisms of in situ tissue regeneration. Here we describe a protocol using a tissue engineering strategy and orthotopic transplantation of either natural decellularized donor tracheae or artificial electrospun nanofiber scaffolds into a rat model. The protocol includes details regarding how to assess the scaffolds' biomechanical properties and cell viability before implantation. It is a reliable and reproducible model that can be used to investigate the crucial aspects and pathways of in situ tracheal tissue restoration and regeneration. The model can be established in
- Published
- 2014
27. RETRACTED: Biomechanical and biocompatibility characteristics of electrospun polymeric tracheal scaffolds
- Author
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Paolo Macchiarini, Silvia Baiguera, Alessandra Bianco, Costantino Del Gaudio, Sebastian Sjöqvist, Greg Lemon, Philipp Jungebluth, Fatemeh Ajalloueian, Ylva Gustafsson, Mei Ling Lim, Antonio Beltrán-Rodríguez, and Johannes C. Haag
- Subjects
Male ,Scaffold ,Materials science ,Biocompatibility ,Polymers ,Polyurethanes ,Biophysics ,Biocompatible Materials ,Cell Count ,Bioengineering ,Rats, Sprague-Dawley ,Biomaterials ,chemistry.chemical_compound ,Bioreactors ,Orthotopic transplantation ,Tissue engineering ,In vivo ,Cell Adhesion ,Polyethylene terephthalate ,Animals ,Tissue Engineering ,Tissue Scaffolds ,Polyethylene Terephthalates ,Mesenchymal Stem Cells ,Electrospinning ,Rats ,Trachea ,Transplantation ,chemistry ,Mechanics of Materials ,Microscopy, Electron, Scanning ,Ceramics and Composites ,Biomedical engineering - Abstract
The development of tracheal scaffolds fabricated based on electrospinning technique by applying different ratios of polyethylene terephthalate (PET) and polyurethane (PU) is introduced here. Prior to clinical implantation, evaluations of biomechanical and morphological properties, as well as biocompatibility and cell adhesion verifications are required and extensively performed on each scaffold type. However, the need for bioreactors and large cell numbers may delay the verification process during the early assessment phase. Hence, we investigated the feasibility of performing biocompatibility verification using static instead of dynamic culture. We performed bioreactor seeding on 3-dimensional (3-D) tracheal scaffolds (PET/PU and PET) and correlated the quantitative and qualitative results with 2-dimensional (2-D) sheets seeded under static conditions. We found that an 8-fold reduction for 2-D static seeding density can essentially provide validation on the qualitative and quantitative evaluations for 3-D scaffolds. In vitro studies revealed that there was notably better cell attachment on PET sheets/scaffolds than with the polyblend. However, the in vivo outcomes of cell seeded PET/PU and PET scaffolds in an orthotopic transplantation model in rodents were similar. They showed that both the scaffold types satisfied biocompatibility requirements and integrated well with the adjacent tissue without any observation of necrosis within 30 days of implantation.
- Published
- 2014
28. Editorial Expression of Concern: Experimental orthotopic transplantation of a tissue-engineered oesophagus in rats
- Author
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Paolo Macchiarini, Ylva Gustafsson, Silvia Baiguera, Cristian Ibarra, Domenico Ribatti, Greg Lemon, Miguel Angel Burguillos, Alessandra Bianco, Sebastian Sjöqvist, Karolina Kublickiene, Heike Kielstein, Bertrand Joseph, Peter Damberg, Costantino Del Gaudio, Philipp Jungebluth, Alexander Sotnichenko, Doris A. Taylor, Rainer Heuchel, Ying Zhao, Johannes C. Haag, Henrik Ullman, Antonio B. Rodriguez, and Mei Ling Lim
- Subjects
0301 basic medicine ,Multidisciplinary ,Tissue engineered ,business.industry ,Science ,digestive, oral, and skin physiology ,General Physics and Astronomy ,General Chemistry ,digestive system ,digestive system diseases ,General Biochemistry, Genetics and Molecular Biology ,Addendum ,03 medical and health sciences ,surgical procedures, operative ,030104 developmental biology ,Orthotopic transplantation ,otorhinolaryngologic diseases ,Cancer research ,Medicine ,business - Abstract
Editorial Expression of Concern: Experimental orthotopic transplantation of a tissue-engineered oesophagus in rats
- Published
- 2016
29. (未来医学研究会のいま<特集III>)栁原操再生医療研究支援寄附金の平成29年度助成活動について Crossing borders in regenerative medicine
- Author
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Sebastian, Sjöqvist
- Published
- 2018
30. Retraction notice to: 'Biomechanical and biocompatibility characteristics of electrospun polymeric tracheal scaffolds '[BIOMATERIALS 35/20 (2014) 5307-5315]
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Antonio Beltrán-Rodríguez, Costantino Del Gaudio, Silvia Baiguera, Alessandra Bianco, Greg Lemon, Ylva Gustafsson, Johannes C. Haag, Sebastian Sjöqvist, Paolo Macchiarini, Philipp Jungebluth, Mei Ling Lim, and Fatemeh Ajalloueian
- Subjects
Biomaterials ,Materials science ,Biocompatibility ,Notice ,Mechanics of Materials ,Biophysics ,Ceramics and Composites ,Bioengineering ,Biomedical engineering - Published
- 2019
31. Decellularized Feeders: An Optimized Method for Culturing Pluripotent Cells
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Paolo Macchiarini, Hero Nikdin, Ivan Vassliev, Christian Unger, Mei Ling Lim, Philipp Jungebluth, Sebastian Sjöqvist, and Kristín Rós Kjartansdóttir
- Subjects
Pluripotent Stem Cells ,KOSR ,Cellular differentiation ,Embryoid body ,Biology ,Cell Line ,Surface-Active Agents ,Cell Adhesion ,Humans ,Enabling Technologies for Cell-Based Clinical Translation ,Induced pluripotent stem cell ,Cell Shape ,Cell potency ,Cell Proliferation ,Homeodomain Proteins ,Extracellular Matrix Proteins ,Induced stem cells ,Feeder Cells ,Gene Expression Regulation, Developmental ,Sodium Dodecyl Sulfate ,Cell Differentiation ,DNA ,Nanog Homeobox Protein ,Cell Biology ,General Medicine ,Fibroblasts ,Embryonic stem cell ,Molecular biology ,Coculture Techniques ,Extracellular Matrix ,Stem cell ,Octamer Transcription Factor-3 ,Biomarkers ,Developmental Biology - Abstract
Pluripotent cells such as human embryonic stem cells and human induced pluripotent stem cells are useful in the field of regenerative medicine because they can proliferate indefinitely and differentiate into all cell types. However, a limiting factor for maintaining and propagating stem cells is the need for inactivated fibroblasts as a growth matrix, since these may potentially cause cross-contamination. In this study, we aimed to maintain stem cells on the extracellular matrix (ECM) of either nonirradiated or γ-irradiated fibroblasts. It has been demonstrated that the ECM contains factors and proteins vital for the adhesion, proliferation, and differentiation of pluripotent cells. In order to preserve the ECM, the cell layers of the fibroblasts were decellularized by treatment with 0.05% sodium dodecyl sulfate (SDS), which resulted in an absence of DNA as compared with conventional feeder culture. However, SDS treatment did not cause a detectable change in the ECM architecture and integrity. Furthermore, immunohistochemistry demonstrated that expressions of major ECM proteins, such as fibronectin, collagen, and laminin, remained unaltered. The human pluripotent cells cultured on this decellularized matrix maintained gene expression of the pluripotency markers NANOG and OCT4 and had the potency to differentiate to three germ layers. The in vitro culture system shown here has an excellent potential since the main allogeneic components (i.e., DNA of the feeder cells) are removed. It is also a technically easy, fast, safe, and cheap method for maintaining a refined feeder-free stem cell culture for further cell differentiation studies.
- Published
- 2013
32. RETRACTED: Verification of cell viability in bioengineered tissues and organs before clinical transplantation
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Philipp Jungebluth, Matthias Corbascio, Silvia Baiguera, Sebastian Sjöqvist, I. V. Gilevich, Oscar E. Simonson, Staffan Strömblad, Paolo Macchiarini, Greg Lemon, Costantino Del Gaudio, Ylva Gustafsson, Mei Ling Lim, Johannes C. Haag, Karl H. Grinnemo, and Fatemeh Ajalloueian
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Cell Survival ,Cell seeding ,Polyurethanes ,Biophysics ,Cell Count ,Bioengineering ,Biology ,Rats, Sprague-Dawley ,Translational Research, Biomedical ,Biomaterials ,Young Adult ,chemistry.chemical_compound ,Cell Adhesion ,medicine ,Animals ,Humans ,DAPI ,Viability assay ,Engineered tissue ,Synthetic scaffold ,Transplantation ,Bioartificial Organs ,Tissue Scaffolds ,Reproducibility of Results ,Mesenchymal Stem Cells ,Rats ,Trachea ,chemistry ,Mechanics of Materials ,Microscopy, Electron, Scanning ,Ceramics and Composites ,Female ,Colorimetric Cell Viability Assay ,Biomedical engineering - Abstract
The clinical outcome of transplantations of bioartificial tissues and organs depends on the presence of living cells. There are still no standard operative protocols that are simple, fast and reliable for confirming the presence of viable cells on bioartificial scaffolds prior to transplantation. By using mathematical modeling, we have developed a colorimetric-based system (colorimetric scale bar) to predict the cell viability and density for sufficient surface coverage. First, we refined a method which can provide information about cell viability and numbers in an in vitro setting: i) immunohistological staining by Phalloidin/DAPI and ii) a modified colorimetric cell viability assay. These laboratory-based methods and the developed colorimetric-based system were then validated in rat transplantation studies of unseeded and seeded tracheal grafts. This was done to provide critical information on whether the graft would be suitable for transplantation or if additional cell seeding was necessary. The potential clinical impact of the colorimetric scale bar was confirmed using patient samples. In conclusion, we have developed a robust, fast and reproducible colorimetric tool that can verify and warrant viability and integrity of an engineered tissue/organ prior to transplantation. This should facilitate a successful transplantation outcome and ensure patient safety.
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- 2013
33. Quantitative uptake of colloidal particles by cell cultures
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Alaa Hassan Said, Sumaira Ashraf, Sathi Roy, Alberto Escudero, Indranath Chakraborty, Daniel Valdeperez, Beatriz Pelaz, Miguel A. Correa Duarte, Neus Feliu, Mei Ling Lim, Atif Masood, Elena Gonzalez, Wolfgang J. Parak, Jonas Hühn, Sebastian Sjöqvist, Philipp Jungebluth, Mikhail V. Zyuzin, European Commission, Lars Hierta Memorial Foundation, Alexander von Humboldt Foundation, Junta de Andalucía, and Ministry of Education (Egypt)
- Subjects
endocrine system ,Environmental Engineering ,Endosome ,Surface Properties ,media_common.quotation_subject ,Cell Culture Techniques ,Nanoparticle ,Uptake ,Nanotechnology ,02 engineering and technology ,010402 general chemistry ,Nanosafety ,01 natural sciences ,complex mixtures ,Cell membrane ,Cellular internalization ,Biological media ,medicine ,Environmental Chemistry ,Humans ,Colloids ,Internalization ,Waste Management and Disposal ,Gold particles ,Cells, Cultured ,media_common ,Life span ,Toxicity ,Chemistry ,Quantum dots ,Cell Membrane ,digestive, oral, and skin physiology ,021001 nanoscience & nanotechnology ,Pollution ,Endocytosis ,0104 chemical sciences ,body regions ,medicine.anatomical_structure ,Colloidal particle ,Cell culture ,Nanoparticles ,0210 nano-technology - Abstract
The use of nanotechnologies involving nano- and microparticles has increased tremendously in the recent past. There are various beneficial characteristics that make particles attractive for a wide range of technologies. However, colloidal particles on the other hand can potentially be harmful for humans and environment. Today, complete understanding of the interaction of colloidal particles with biological systems still remains a challenge. Indeed, their uptake, effects, and final cell cycle including their life span fate and degradation in biological systems are not fully understood. This is mainly due to the complexity of multiple parameters which need to be taken in consideration to perform the nanosafety research. Therefore, we will provide an overview of the common denominators and ideas to achieve universal metrics to assess their safety. The review discusses aspects including how biological media could change the physicochemical properties of colloids, how colloids are endocytosed by cells, how to distinguish between internalized versus membrane-attached colloids, possible correlation of cellular uptake of colloids with their physicochemical properties, and how the colloidal stability of colloids may vary upon cell internalization. In conclusion three main statements are given. First, in typically exposure scenarios only part of the colloids associated with cells are internalized while a significant part remain outside cells attached to their membrane. For quantitative uptake studies false positive counts in the form of only adherent but not internalized colloids have to be avoided. pH sensitive fluorophores attached to the colloids, which can discriminate between acidic endosomal/lysosomal and neutral extracellular environment around colloids offer a possible solution. Second, the metrics selected for uptake studies is of utmost importance. Counting the internalized colloids by number or by volume may lead to significantly different results. Third, colloids may change their physicochemical properties along their life cycle, and appropriate characterization is required during the different stages., This work was supported by the European Commission (grant FutureNanoNeeds) grant agreement no. 604602 to WJP. NF acknowledges funding from the Lars Hiertas Minne Fundation (Sweden), SA, BP and IC acknowledge a fellowship from the Alexander von Humboldt Fundation (Germany). AE acknowledges Junta de Andalucía (Spain) for a Talentia Postdoc Fellowship, co-financed by the European Union Seventh Framework Programme, grant agreement no 267226. AHS acknowledges the Egyptian government (Ministry of Higher Education, Mission). The project was also supported by the Dr. Dorka-Stiftung (Germany) to PJ.
- Published
- 2016
34. ORAI1-mediated calcium influx is required for human cytotoxic lymphocyte degranulation and target cell lysis
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Stephan Ehl, Anne Rensing-Ehl, Yenan T. Bryceson, Christie-Ann McCarl, Birthe Jessen, Sebastian Fuchs, Stefan Feske, Markus Hufnagel, Hanspeter Pircher, Stephanie M. Wood, Cyril Fauriat, Andrea Maul-Pavicic, Klaus Schwarz, Katsuhiko Mikoshiba, Wolfgang W. A. Schamel, Samuel C. C. Chiang, Ilka Schulze, Thilo Bass, and Sebastian Sjöqvist
- Subjects
Cytotoxicity, Immunologic ,ORAI1 Protein ,Cell Degranulation ,medicine.medical_treatment ,Lymphocyte ,Biology ,Interferon-gamma ,Interleukin 21 ,medicine ,Humans ,Cytotoxic T cell ,Chemokine CCL4 ,Multidisciplinary ,Lymphokine-activated killer cell ,Tumor Necrosis Factor-alpha ,Interleukins ,Degranulation ,Biological Sciences ,Cell biology ,Killer Cells, Natural ,Cytokine ,medicine.anatomical_structure ,Perforin ,biology.protein ,Cytokines ,Calcium ,Calcium Channels ,T-Lymphocytes, Cytotoxic - Abstract
Lymphocytes mediate cytotoxicity by polarized release of the contents of cytotoxic granules toward their target cells. Here, we have studied the role of the calcium release-activated calcium channel ORAI1 in human lymphocyte cytotoxicity. Natural killer (NK) cells obtained from an ORAI1-deficient patient displayed defective store-operated Ca 2+ entry (SOCE) and severely defective cytotoxic granule exocytosis leading to impaired target cell lysis. Similar findings were obtained using NK cells from a stromal interaction molecule 1-deficient patient. The defect occurred at a late stage of the signaling process, because activation of leukocyte functional antigen (LFA)-1 and cytotoxic granule polarization were not impaired. Moreover, pharmacological inhibition of SOCE interfered with degranulation and target cell lysis by freshly isolated NK cells and CD8 + effector T cells from healthy donors. In addition to effects on lymphocyte cytotoxicity, synthesis of the chemokine macrophage inflammatory protein-1β and the cytokines TNF-α and IFN-γ on target cell recognition was impaired in ORAI1-deficient NK cells, as previously described for T cells. By contrast, NK cell cytokine production induced by combinations of IL-12, IL-15, and IL-18 was not impaired by ORAI1 deficiency. Taken together, these results identify a critical role for ORAI1-mediated Ca 2+ influx in granule exocytosis for lymphocyte cytotoxicity as well as for cytokine production induced by target cell recognition.
- Published
- 2011
35. Intratracheale Anwendung von autologen Stammzellen in Patienten mit ARDS
- Author
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Matthew J.A. Wood, H Kalzén, Tom Luedde, B Holzgraefe, H Dienemann, Evren Alici, Ola Hermanson, S El Andaloussi, Ana I. Teixeira, Oscar P. B. Wiklander, Ola Winqvist, Joel Nordin, Philipp Jungebluth, Petra Jones, Vanessa Lundin, Sebastian Sjöqvist, M.L. Lim, and Adil Doganay Duru
- Subjects
Gynecology ,medicine.medical_specialty ,business.industry ,Medicine ,Surgery ,business - Published
- 2015
36. Orthotopic transplantation of a tissue engineered diaphragm in rats
- Author
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Philipp Jungebluth, S. S. Dzhimak, Geoanna Bautista, Konstantin A. Danilenko, Sergei N. Chvalun, I. S. Gumenyuk, I. V. Gilevich, Mei Ling Lim, Paolo Macchiarini, Mark J. Holterman, E. A. Gubareva, Timofei E. Grigoriev, Greg Lemon, Doris A. Taylor, Antonio Beltrán Rodríguez, Sebastian Sjöqvist, E. V. Kuevda, A. S. Sotnichenko, Alexander G. Pokhotko, Ylva Gustafsson, Vladimir M. Pokrovsky, R. Z. Nakokhov, Neus Feliu, A. A. Basov, and S. V. Krasheninnikov
- Subjects
0301 basic medicine ,Male ,Pathology ,medicine.medical_specialty ,Transplantation, Heterotopic ,Diaphragm ,Biophysics ,Diaphragmatic breathing ,Neovascularization, Physiologic ,Transplants ,Bioengineering ,Mesenchymal Stem Cell Transplantation ,Biomaterials ,03 medical and health sciences ,0302 clinical medicine ,Bioreactors ,Tissue engineering ,Absorbable Implants ,medicine ,Cell Adhesion ,Animals ,Wound Healing ,Decellularization ,Tissue Engineering ,Tissue Scaffolds ,business.industry ,Electromyography ,Regeneration (biology) ,Macrophages ,Mesenchymal stem cell ,Graft Survival ,Cell Differentiation ,Allografts ,Diaphragm (structural system) ,Rats ,Transplantation ,Radiography ,030104 developmental biology ,medicine.anatomical_structure ,Mechanics of Materials ,Rats, Inbred Lew ,030220 oncology & carcinogenesis ,Ceramics and Composites ,Bone marrow ,business ,Hernias, Diaphragmatic, Congenital - Abstract
The currently available surgical options to repair the diaphragm are associated with significant risks of defect recurrence, lack of growth potential and restored functionality. A tissue engineered diaphragm has the potential to improve surgical outcomes for patients with congenital or acquired disorders. Here we show that decellularized diaphragmatic tissue reseeded with bone marrow mesenchymal stromal cells (BM-MSCs) facilitates in situ regeneration of functional tissue. A novel bioreactor, using simultaneous perfusion and agitation, was used to rapidly decellularize rat diaphragms. The scaffolds retained architecture and mechanical properties and supported cell adhesion, proliferation and differentiation. Biocompatibility was further confirmed in vitro and in vivo. We replaced 80% of the left hemidiaphragm with reseeded diaphragmatic scaffolds. After three weeks, transplanted animals gained 32% weight, showed myography, spirometry parameters, and histological evaluations similar to native rats. In conclusion, our study suggested that reseeded decellularized diaphragmatic tissue appears to be a promising option for patients in need of diaphragmatic reconstruction.
- Published
- 2015
37. Autologous Peripheral Blood Mononuclear Cells as Treatment in Refractory Acute Respiratory Distress Syndrome
- Author
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B. Anderstam, Philipp Jungebluth, Ola Winqvist, S El Andaloussi, J.C. Haag, Michael Chrobok, A. Beltrán Rodríguez, Ola Hermanson, H Kalzén, Wiklander Opb., Evren Alici, K.G. Roddewig, Christoph Roderburg, M.L. Lim, Vanessa Lundin, Tom Luedde, Ylva Gustafsson, Nina Heldring, Ana I. Teixeira, P Macchiarini, Adil Doganay Duru, Sebastian Sjöqvist, Wood Mja., Petra Jones, Joel Nordin, and B Holzgraefe
- Subjects
Pulmonary and Respiratory Medicine ,Male ,ARDS ,medicine.medical_treatment ,Multiple Organ Failure ,Down-Regulation ,Inflammation ,Peripheral blood mononuclear cell ,Transplantation, Autologous ,Cell therapy ,Young Adult ,Extracorporeal Membrane Oxygenation ,Fatal Outcome ,medicine ,Extracorporeal membrane oxygenation ,Humans ,Respiratory function ,Erythropoietin ,Respiratory Distress Syndrome ,business.industry ,medicine.disease ,Cadherins ,Up-Regulation ,Transplantation ,MicroRNAs ,Immunology ,Leukocytes, Mononuclear ,Cytokines ,Snail Family Transcription Factors ,medicine.symptom ,business ,medicine.drug ,Transcription Factors - Abstract
Background: Acute respiratory distress syndrome (ARDS) is a devastating disorder. Despite enormous efforts in clinical research, effective treatment options are lacking, and mortality rates remain unacceptably high. Objectives: A male patient with severe ARDS showed no clinical improvement with conventional therapies. Hence, an emergent experimental intervention was performed. Methods: We performed intratracheal administration of autologous peripheral blood-derived mononuclear cells (PBMCs) and erythropoietin (EPO). Results: We found that after 2 days of initial PBMC/EPO application, lung function improved and extracorporeal membrane oxygenation (ECMO) support was reduced. Bronchoscopy and serum inflammatory markers revealed reduced inflammation. Additionally, serum concentration of miR-449a, b, c and miR-34a, a transient upregulation of E-cadherin and associated chromatin marks in PBMCs indicated airway epithelial differentiation. Extracellular vesicles from PBMCs demonstrated anti-inflammatory capacity in a TNF-a-mediated nuclear factor-κB in vitro assay. Despite improving respiratory function, the patient died of multisystem organ failure on day 38 of ECMO treatment. Conclusions: This case report provides initial encouraging evidence to use locally instilled PBMC/EPO for treatment of severe refractory ARDS. The observed clinical improvement may partially be due to the anti-inflammatory effects of PBMC/EPO to promote tissue regeneration. Further studies are needed for more in-depth understanding of the underlying mechanisms of in vivo regeneration.
- Published
- 2015
38. Tissue Engineering of the Esophagus
- Author
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Paolo Macchiarini and Sebastian Sjöqvist
- Subjects
medicine.medical_specialty ,business.industry ,Stomach ,Mucosal cell ,Cancer ,Esophageal Disorder ,medicine.disease ,Regenerative medicine ,Surgery ,medicine.anatomical_structure ,Tissue engineering ,Small animal ,Medicine ,Esophagus ,business - Abstract
Tissue engineering has the potential to help many adults and children affected by esophageal disorders such as cancer, Barrett’s esophagus, malformations, trauma, etc. Current surgical treatment is a complex procedure where the esophagus is replaced with either colon or stomach. Related mortality and morbidity is high, and functional outcomes are poor. A tissue-engineered graft could eliminate the need to use the patients’ own tissue and could result in a better clinical outcome. Tissue engineering approaches are also used to facilitate mucosal and submucosal healing after endoscopic removal of superficial cancer or precancerous lesions. This chapter describes the anatomy, physiology, and pathology of the esophagus, and summarizes the efforts made so far in the world of tissue engineering of the esophagus. We describe the use of biological and synthetic scaffolds and discuss results from many large and small animal publications. Lastly, we present two clinical studies that have been published to date, where either extra cellular matrix or oral mucosal cell sheets were transplanted to the wound bed after interventional endoscopy. In conclusion, the field of regenerative medicine and tissue engineering has the potential to save many lives, but much effort still needs to be made.
- Published
- 2015
39. List of Contributors
- Author
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Samad Ahadian, Kentaro Akiyaman, Sarah Alansari, Mani Alikhani, Mona Alikhani, Agnieszka Arthur, Shylaja Arulkumar, Silvia Baiguera, Alessandra Bianco, Nabil F. Bissada, Françoise Bleicher, Jacqueline M. Bliley, Dieter D. Bosshardt, Tatiana M. Botero, Raquel Braga, Kristen K. Briggs, Patricia J. Brooks, Kevin Cannon, Florence Carrouel, Miquella G. Chavez, Ming-Te Cheng, George J. Christ, Colin A. Cook, Paul R. Cooper, Timothy C. Cox, Michael L. Cunningham, Jesse Dashe, Ze’ev Davidovitch, Lindsay C. Davies, Costantino Del Gaudio, Thomas G.H. Diekwisch, Anibal Diogenes, Randall L. Duncan, Paul C. Edwards, Tarek El-Bialy, Donald H. Enlow, Mary C. Farach-Carson, Jean-Christophe Farges, Stephen E. Feinberg, Hady Felfy, Phillip N. Freeman, Martin Freilich, Toshinori Fujie, Changyou Gao, William V. Giannobile, Siew-Ging Gong, Warren L. Grayson, Robert M. Greene, Stan Gronthos, Reinhard Gruber, Lari Häkkinen, Craig Hanna, Kenneth M. Hargreaves, Emilio Hara Satoshi, Daniel A. Harrington, Basma Hashmi, Jyrki Heino, Anne V. Hing, Christina Holmes, Masaki J. Honda, Jeremy A. Horst, Michael S. Hu, Francis J. Hughes, Ben P. Hung, Pinar Yilgor Huri, Donald E. Ingber, Keitaro Isokawa, Kenji Izumi, Eric N. James, Xinqiao Jia, Yan Jin, E. Emily Joo, Emma Juuri, Hideaki Kagami, Hirokazu Kaji, Mo K. Kang, A. Kashi, Hiroko Kato, Jasvir Kaur, Letitia V. Keller, Paul D. Kemp, Ali Khademhosseini, Edmund Khoo, Hae-Won Kim, William J. King, Richard E. Kirschner, Ophir D. Klein, Jonathan C. Knowles, Leeni Koivisto, Tunay Kökten, Paul H. Krebsbach, Takuo Kuboki, Sabine Kuchler-Bopp, Simone Kucska, Liisa Kuhn, Hannu Larjava, Eun-Jung Lee, Joo-Hee Lee, Oscar K. Lee, Yoo Bin Lee, Hervé Lesot, Mark P. Lewis, Bei Li, Luyan Li, Jung Yul Lim, Xing Liu, Xiaohua Liu, David D. Lo, Michael T. Longaker, H. Peter Lorenz, David G. Lott, Vlasta Lungová, Lie Ma, Paolo Macchiarini, Tadanori Mammoto, Mona K. Marei, Darja Marolt, Kacey G. Marra, Neil R.W. Martin, Eva Matalová, Tomokazu Matsue, Shebli Mehrazarin, Mina Mina, Michel Modo, Jaroslav Mokry, Christian Morsczeck, Rania M. Moussa, Neal C. Murphy, Vagan Mushegyan, Naglaa B. Nagy, Lakshmi S. Nair, Harry Nayar, Jacques E. Nör, Raquel Obregón, Tae-Ju Oh, Kyoko Oka, Serge Ostrovidov, No-Hee Park, Michael J. Passineau, Juliana A. Passipieri, Rishikaysh Pisal, Darren Player, Swati Pradhan-Bhatt, Selvakumar Prakash Parthiban, Jan Prochazka, Michaela Prochazkova, Tiejun Qu, Murugan Ramalingam, Javier Ramón-Azcón, Deepti Rana, Herbert L. Ray, David A. Reed, Dieter P. Reinhardt, Béatrice Richard, Brandon D. Riehl, Hector F. Rios, J. Peter Rubin, Manal M. Saad, Ashneet Sachar, Robert L. Sah, Subrata Saha, Yoko Saito, Kaarunya Sampathkumar, Chinapa Sangsuwon, Byoung Moo Seo, Paul Sharpe, Songtao Shi, Yu-Ru V. Shih, Hitoshi Shiku, Seung Yun Shin, Sebastian Sjöqvist, Alastair J. Sloan, Anthony (Tony) J. Smith, In Seok Song, Neil Stephens, Phil Stephens, Kathy K.H. Svoboda, Maryam Tabrizian, Cristina Teixeira, Fabricio B. Teixeira, Joshua P. Temple, Irma Thesleff, Béatrice Thivichon-Prince, Taku Toriumi, Trevor E. Treasure, Soyoun Um, Ajaykumar Vishwakarma, Chunfen Wang, Deborah Watson, Jeffrey B. Watson, Bo Wen, Robert L. Witt, Mark E. Wong, Kenneth M. Yamada, Hala H. Yassin, Daniel Zakheim, Bing Zhang, and Andrew S. Zimmermann
- Published
- 2015
40. Characterization of Stem-Like Cells in Mucoepidermoid Tracheal Paediatric Tumor
- Author
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Jianri Lim, Paolo Macchiarini, Silvia Baiguera, Antonio Beltrán Rodríguez, Greg Lemon, Brandon Nick Sern Ooi, Tom Luedde, Lars Ährlund-Richter, Magnus Nordenskjöld, Evren Alici, Ivan Vassiliev, Ylva Gustafsson, Agne Liedén, José Inzunza, Iyadh Douagi, Johannes C. Haag, Philipp Jungebluth, Duncan Baker, Alina Popova, I. V. Gilevich, Sebastian Sjöqvist, Christian Unger, Mei Ling Lim, Isabell Hultman, and Jurate Asmundsson
- Subjects
Male ,Pathology ,Microarrays ,Cellular differentiation ,lcsh:Medicine ,Cell Separation ,Cell Fate Determination ,Pediatrics ,Mice ,Animal Cells ,Molecular Cell Biology ,Medicine and Health Sciences ,lcsh:Science ,Child ,Stem cell transplantation for articular cartilage repair ,Multidisciplinary ,Stem Cells ,Amniotic stem cells ,Cell Differentiation ,Genomics ,3. Good health ,Mucoepidermoid Tumor ,medicine.anatomical_structure ,Bioassays and Physiological Analysis ,Oncology ,Neoplastic Stem Cells ,Female ,Stem cell ,Cellular Types ,Research Article ,medicine.medical_specialty ,Research and Analysis Methods ,Cancer stem cell ,medicine ,Animals ,Humans ,business.industry ,Gene Expression Profiling ,Mesenchymal stem cell ,lcsh:R ,Biology and Life Sciences ,Computational Biology ,Mesenchymal Stem Cells ,Cell Biology ,Pediatric Oncology ,lcsh:Q ,Tracheal Neoplasms ,Bone marrow ,business ,Developmental Biology - Abstract
Stem cells contribute to regeneration of tissues and organs. Cells with stem cell-like properties have been identified in tumors from a variety of origins, but to our knowledge there are yet no reports on tumor-related stem cells in the human upper respiratory tract. In the present study, we show that a tracheal mucoepidermoid tumor biopsy obtained from a 6 year-old patient contained a subpopulation of cells with morphology, clonogenicity and surface markers that overlapped with bone marrow mesenchymal stromal cells (BM-MSCs). These cells, designated as MEi (mesenchymal stem cell-like mucoepidermoid tumor) cells, could be differentiated towards mesenchymal lineages both with and without induction, and formed spheroids in vitro. The MEi cells shared several multipotent characteristics with BM-MSCs. However, they displayed differences to BM-MSCs in growth kinectics and gene expression profiles relating to cancer pathways and tube development. Despite this, the MEi cells did not possess in vivo tumor-initiating capacity, as proven by the absence of growth in situ after localized injection in immunocompromised mice. Our results provide an initial characterization of benign tracheal cancer-derived niche cells. We believe that this report could be of importance to further understand tracheal cancer initiation and progression as well as therapeutic development.
- Published
- 2014
41. Experimental orthotopic transplantation of a tissue-engineered oesophagus in rats
- Author
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Alessandra Bianco, Greg Lemon, Henrik Ullman, Alexander Sotnichenko, Silvia Baiguera, Karolina Kublickiene, Cristian Ibarra, Antonio Beltrán Rodríguez, Rainer Heuchel, Costantino Del Gaudio, Paolo Macchiarini, Ying Zhao, Philipp Jungebluth, Miguel Angel Burguillos, Sebastian Sjöqvist, Heike Kielstein, Bertrand Joseph, Domenico Ribatti, Johannes C. Haag, Doris A. Taylor, Peter Damberg, Ylva Gustafsson, and Mei Ling Lim
- Subjects
Pathology ,medicine.medical_specialty ,Angiogenesis ,Settore ING-IND/22 - Scienza e Tecnologia dei Materiali ,Cellular differentiation ,General Physics and Astronomy ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Esophagus ,0302 clinical medicine ,Tissue engineering ,medicine ,Animals ,Myocyte ,030304 developmental biology ,0303 health sciences ,Multidisciplinary ,Tissue Engineering ,Tissue Scaffolds ,Regeneration (biology) ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,General Chemistry ,Anatomy ,Epithelium ,medicine.anatomical_structure ,030220 oncology & carcinogenesis - Abstract
A tissue-engineered oesophageal scaffold could be very useful for the treatment of pediatric and adult patients with benign or malignant diseases such as carcinomas, trauma or congenital malformations. Here we decellularize rat oesophagi inside a perfusion bioreactor to create biocompatible biological rat scaffolds that mimic native architecture, resist mechanical stress and induce angiogenesis. Seeded allogeneic mesenchymal stromal cells spontaneously differentiate (proven by gene-, protein and functional evaluations) into epithelial- and muscle-like cells. The reseeded scaffolds are used to orthotopically replace the entire cervical oesophagus in immunocompetent rats. All animals survive the 14-day study period, with patent and functional grafts, and gain significantly more weight than sham-operated animals. Explanted grafts show regeneration of all the major cell and tissue components of the oesophagus including functional epithelium, muscle fibres, nerves and vasculature. We consider the presented tissue-engineered oesophageal scaffolds a significant step towards the clinical application of bioengineered oesophagi., Patients with oesophageal diseases may require surgical removal and replacement of the oesophagus. Here the authors seed mesenchymal stromal cells on a decellularized rat oesophagus and show that this bioengineered tissue construct restores swallowing function after transplantation into rats.
- Published
- 2014
42. Preservation of aortic root architecture and properties using a detergent-enzymatic perfusion protocol
- Author
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Ana I. Teixeira, Greg Lemon, Bertrand Joseph, Paolo Macchiarini, Philipp Jungebluth, Johannes C. Haag, Alexander Sotnichenko, Ylva Gustafsson, Vanessa Lundin, Fatemeh Ajalloueian, Mei Ling Lim, Linda Helen Friedrich, Heike Kielstein, Miguel Angel Burguillos, and Sebastian Sjöqvist
- Subjects
Aortic valve ,medicine.medical_specialty ,Materials science ,Biocompatibility ,Cell Survival ,medicine.medical_treatment ,Detergents ,Biophysics ,Bioengineering ,Biomaterials ,Valve replacement ,Tissue engineering ,medicine ,Cell Adhesion ,Animals ,Cells, Cultured ,Decellularization ,Tissue Engineering ,Tissue Processing ,Mesenchymal Stem Cells ,Immunohistochemistry ,Surgery ,Tissue Degeneration ,medicine.anatomical_structure ,Mechanics of Materials ,Aortic Valve ,Ceramics and Composites ,Cell activation ,Biomedical engineering - Abstract
Aortic valve degeneration and dysfunction is one of the leading causes for morbidity and mortality. The conventional heart-valve prostheses have significant limitations with either life-long anticoagulation therapeutic associated bleeding complications (mechanical valves) or limited durability (biological valves). Tissue engineered valve replacement recently showed encouraging results, but the unpredictable outcome of tissue degeneration is likely associated to the extensive tissue processing methods. We believe that optimized decellularization procedures may provide aortic valve/root grafts improved durability. We present an improved/innovative decellularization approach using a detergent-enzymatic perfusion method, which is both quicker and has less exposure of matrix degenerating detergents, compared to previous protocols. The obtained graft was characterized for its architecture, extracellular matrix proteins, mechanical and immunological properties. We further analyzed the engineered aortic root for biocompatibility by cell adhesion and viability in vitro and heterotopic implantation in vivo. The developed decellularization protocol was substantially reduced in processing time whilst maintaining tissue integrity. Furthermore, the decellularized aortic root remained bioactive without eliciting any adverse immunological reaction. Cell adhesion and viability demonstrated the scaffold's biocompatibility. Our optimized decellularization protocol may be useful to develop the next generation of clinical valve prosthesis with a focus on improved mechanical properties and durability.
- Published
- 2013
43. Whole organ and tissue reconstruction in thoracic regenerative surgery
- Author
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Paolo Macchiarini, Linda Helen Friedrich, Greg Lemon, Fatemeh Ajalloueian, I. V. Gilevich, Karl-Henrik Grinnemo, Arthur L. Caplan, Philipp Jungebluth, Mei Ling Lim, J.C. Haag, E. A. Gubareva, and Sebastian Sjöqvist
- Subjects
Pathology ,medicine.medical_specialty ,Tissue replacement ,MEDLINE ,Tissue reconstruction ,030204 cardiovascular system & hematology ,Biology ,Regenerative Medicine ,Organ transplantation ,03 medical and health sciences ,Immune System Phenomena ,0302 clinical medicine ,Bioreactors ,Tissue engineering ,medicine ,Humans ,Cardiac Surgical Procedures ,Intensive care medicine ,Adverse effect ,Lung ,Digestive System Surgical Procedures ,030304 developmental biology ,0303 health sciences ,Tissue Engineering ,Tissue Scaffolds ,Respiratory dysfunction ,General Medicine ,Organ Transplantation ,Thoracic Surgical Procedures ,3. Good health ,Transplantation ,Trachea ,Larynx ,Stem Cell Transplantation - Abstract
Development of novel prognostic, diagnostic, and treatment options will provide major benefits for millions of patients with acute or chronic respiratory dysfunction, cardiac-related disorders, esophageal problems, or other diseases in the thorax. Allogeneic organ transplant is currently available. However, it remains a trap because of its dependency on a very limited supply of donated organs, which may be needed for both initial and subsequent transplants. Furthermore, it requires lifelong treatment with immunosuppressants, which are associated with adverse effects. Despite early clinical applications of bioengineered organs and tissues, routine implementation is still far off. For this review, we searched the PubMed, MEDLINE, and Ovid databases for the following keywords for each tissue or organ: tissue engineering, biological and synthetic scaffold/graft, acellular and decelluar(ized), reseeding, bioreactor, tissue replacement, and transplantation. We identified the current state-of-the-art practices in tissue engineering with a focus on advances during the past 5 years. We discuss advantages and disadvantages of biological and synthetic solutions and introduce novel strategies and technologies for the field. The ethical challenges of innovation in this area are also reviewed.
- Published
- 2012
44. Decellularized Lung Approach to Understand Cell-Matrix Cues
- Author
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Paolo Macchiarini, Greg Lemon, Mei Ling Lim, Philipp Jungebluth, Sebastian Sjöqvist, and R. Amin
- Subjects
Cancer Research ,Transplantation ,Decellularization ,Lung ,Chemistry ,Immunology ,Cell Biology ,Cell biology ,Extracellular matrix ,medicine.anatomical_structure ,Oncology ,medicine ,Immunology and Allergy ,Genetics (clinical) - Published
- 2016
45. Individualized Decellularization for Tissue Engineering Tissues and Organs in Animals and Humans
- Author
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Paolo Macchiarini, R. Amin, Sebastian Sjöqvist, Greg Lemon, and Mei Ling Lim
- Subjects
Cancer Research ,Transplantation ,Pathology ,medicine.medical_specialty ,Decellularization ,Immunology ,Cell ,Adipose tissue ,Cell Biology ,Pet imaging ,Anatomy ,Biology ,In vitro ,medicine.anatomical_structure ,Oncology ,Tissue engineering ,medicine ,Immunology and Allergy ,Bone marrow ,Stem cell ,health care economics and organizations ,Genetics (clinical) - Abstract
coding the human or species specific NIS genes. NIS function in transduced MSC was first validated in vitro; NIS expressing MSC (MSC_NIS) from multiple species concentrated high levels of I-125 with no side effects. The sensitivity of cell detection was determined by transplanting a known number of MSC_NIS subcutaneously into mice. We can reliably detect 2x105 MSC_NIS in mice using the newly acquired U-SPECT II machine. Canine MSC derived from the bone marrow were surprisingly robust; viable cells were still detected (albeit lower numbers) at day 28 in the athymic mice. In contrast, NIS signals from adipose tissue derived rat or human MSC disappeared by day 7 post transplantation. Using PET imaging and F18-TFB, sensitivity of imaging could be significantly improved to detect lower numbers of cells. We are currently evaluating the use of NIS for tracking natural killer cells and transplanted hemapoietic stem cells and results will be presented.
- Published
- 2016
46. Modelling biological cell attachment and growth on adherent surfaces
- Author
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Philipp Jungebluth, Sebastian Sjöqvist, Johannes C. Haag, Greg Lemon, Ylva Gustafsson, Paolo Macchiarini, Fatemeh Ajalloueian, and Mei Ling Lim
- Subjects
Materials science ,Thin layers ,Applied Mathematics ,Mesenchymal stem cell ,Nanotechnology ,Mesenchymal Stem Cells ,Numerical Analysis, Computer-Assisted ,Agricultural and Biological Sciences (miscellaneous) ,Models, Biological ,Rats ,chemistry.chemical_compound ,chemistry ,Tissue engineering ,Cell culture ,Modeling and Simulation ,Monolayer ,Polyethylene terephthalate ,Biophysics ,Cell Adhesion ,Deposition (phase transition) ,Animals ,Seeding ,Cell Proliferation - Abstract
A mathematical model, in the form of an integro-partial differential equation, is presented to describe the dynamics of cells being deposited, attaching and growing in the form of a monolayer across an adherent surface. The model takes into account that the cells suspended in the media used for the seeding have a distribution of sizes, and that the attachment of cells restricts further deposition by fragmenting the parts of the domain unoccupied by cells. Once attached the cells are assumed to be able to grow and proliferate over the domain by a process of infilling of the interstitial gaps; it is shown that without cell proliferation there is a slow build up of the monolayer but if the surface is conducive to cell spreading and proliferation then complete coverage of the domain by the monolayer can be achieved more rapidly. Analytical solutions of the model equations are obtained for special cases, and numerical solutions are presented for parameter values derived from experiments of rat mesenchymal stromal cells seeded onto thin layers of collagen-coated polyethylene terephthalate electrospun fibers. The model represents a new approach to describing the deposition, attachment and growth of cells over adherent surfaces, and should prove useful for studying the dynamics of the seeding of biomaterials.
- Published
- 2012
47. Viability and proliferation of rat MSCs on adhesion protein-modified PET and PU scaffolds
- Author
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Alessandra Bianco, Vanessa Lundin, Greg Lemon, Philipp Jungebluth, Ylva Gustafsson, Fatemeh Ajalloueian, Guido Moll, Sebastian Sjöqvist, Mei Ling Lim, Silvia Baiguera, Johannes C. Haag, Paolo Macchiarini, Costantino Del Gaudio, and Ana I. Teixeira
- Subjects
Male ,Materials science ,Cell Survival ,Settore ING-IND/22 - Scienza e Tecnologia dei Materiali ,Polyurethanes ,Biophysics ,Bioengineering ,Biomaterials ,Extracellular matrix ,Coated Materials, Biocompatible ,Tissue engineering ,Cell Adhesion ,Animals ,RNA, Messenger ,Viability assay ,Cell adhesion ,Cells, Cultured ,Cell Proliferation ,Tissue Engineering ,Tissue Scaffolds ,biology ,Polyethylene Terephthalates ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Adhesion ,Extracellular Matrix ,Rats ,Fibronectin ,Transplantation ,Rats, Inbred Lew ,Mechanics of Materials ,Ceramics and Composites ,biology.protein ,Biomedical engineering - Abstract
In 2011, the first in-man successful transplantation of a tissue engineered trachea-bronchial graft, using a synthetic POSS-PCU nanocomposite construct seeded with autologous stem cells, was performed. To further improve this technology, we investigated the feasibility of using polymers with a three dimensional structure more closely mimicking the morphology and size scale of native extracellular matrix (ECM) fibers. We therefore investigated the in vitro biocompatibility of electrospun polyethylene terephthalate (PET) and polyurethane (PU) scaffolds, and determined the effects on cell attachment by conditioning the fibers with adhesion proteins. Rat mesenchymal stromal cells (MSCs) were seeded on either PET or PU fiber-layered culture plates coated with laminin, collagen I, fibronectin, poly-D-lysine or gelatin. Cell density, proliferation, viability, morphology and mRNA expression were evaluated. MSC cultures on PET and PU resulted in similar cell densities and amounts of proliferating cells, with retained MSC phenotype compared to data obtained from tissue culture plate cultures. Coating the scaffolds with adhesion proteins did not increase cell density or cell proliferation. Our data suggest that both PET and PU mats, matching the dimensions of ECM fibers, are biomimetic scaffolds and, because of their high surface area-to-volume provided by the electrospinning procedure, makes them per se suitable for cell attachment and proliferation without any additional coating.
- Published
- 2012
48. Basic Considerations with Cell Sheets
- Author
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Masayuki Yamato and Sebastian Sjöqvist
- Subjects
Extracellular matrix ,Transplantation ,Immune system ,Tissue engineering ,Chemistry ,Biodegradable scaffold ,Esophageal surgery ,Host tissue ,Cell sheet ,Cell biology - Abstract
The foundation of cell sheet engineering is to harvest cells in a minimally destructive way. This way, the cells that retain their cell-to-cell contact show a higher expression of surface proteins, and retain their more highly developed functions than cells harvested enzymatically. In contrast to using biodegradable scaffolds for tissue engineering, cell sheets triggers a significantly lower immune response and do not cause an excess of extracellular matrix to be secreted around the transplanted cells. Cell sheets also tend to integrate more easily with host tissue. In this chapter, the basic concept and history of cell sheet engineering are discussed, including some of our own studies. This chapter focuses on using cell sheets for constructing corneas, minimizing postoperative complications after esophageal surgery, and transplantation of hormone-producing endocrine tissues.
- Published
- 2011
49. Saliva and Saliva Extracellular Vesicles for Biomarker Candidate Identification—Assay Development and Pilot Study in Amyotrophic Lateral Sclerosis
- Author
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Sebastian Sjoqvist and Kentaro Otake
- Subjects
saliva ,extracellular vesicles ,biomarkers ,amyotrophic lateral sclerosis ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Saliva is gaining increasing attention as a source of biomarkers due to non-invasive and undemanding collection access. Extracellular vesicles (EVs) are nano-sized, cell-released particles that contain molecular information about their parent cells. In this study, we developed methods for saliva biomarker candidate identification using EV-isolation and proteomic evaluation. We used pooled saliva samples for assay development. EVs were isolated using membrane affinity-based methods followed by their characterization using nanoparticle tracking analysis and transmission electron microscopy. Subsequently, both saliva and saliva-EVs were successfully analyzed using proximity extension assay and label-free quantitative proteomics. Saliva-EVs had a higher purity than plasma-EVs, based on the expression of EV-proteins and albumin. The developed methods could be used for the analysis of individual saliva samples from amyotrophic lateral sclerosis (ALS) patients and controls (n = 10 each). The starting volume ranged from 2.1 to 4.9 mL and the amount of total isolated EV-proteins ranged from 5.1 to 42.6 µg. Although no proteins were significantly differentially expressed between the two groups, there was a trend for a downregulation of ZNF428 in ALS-saliva-EVs and an upregulation of IGLL1 in ALS saliva. In conclusion, we have developed a robust workflow for saliva and saliva-EV analysis and demonstrated its technical feasibility for biomarker discovery.
- Published
- 2023
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50. Regenerative medicine for esophageal reconstruction after cancer treatment
- Author
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Matthius Löhr, Takeshi Ohki, Makoto Kondo, Pontus Blomberg, Susumu Eguchi, Nobuo Kanai, Masayuki Yamato, and Sebastian Sjöqvist
- Subjects
lcsh:R5-920 ,medicine.medical_specialty ,lcsh:Cytology ,business.industry ,medicine.medical_treatment ,Biomedical Engineering ,MEDLINE ,Regenerative medicine ,law.invention ,Cancer treatment ,Surgery ,Biomaterials ,Clinical trial ,medicine.anatomical_structure ,Randomized controlled trial ,law ,Esophagectomy ,Letter to the editor ,Medicine ,lcsh:QH573-671 ,Esophagus ,Stem cell ,lcsh:Medicine (General) ,business ,Developmental Biology - Abstract
We read with interest a review of the esophageal reconstruction reported by Kerm and colleagues [1], and would like to add some information pertaining to the various therapies covering endosurgical defects. The authors pointed out that few patients were included in the clinical trials and a more extensive clinical study is needed to establish the efficacy. They further state that fullthickness tissue replacement after esophagectomy remains a challenge. Advances in stem cell and cell-sheet technologies could provide the ultimate solution to complications after esophagectomy, e.g. postsurgical strictures and stenoses. We agree. As mentioned in their fine review, we published a pilot study in ten patients with autologous oral mucosal cells cultivated on membranes (cell sheet) [1]. In the meantime, we have performed additional prospective clinical studies in Sweden as well as in Japanwhere 30 patients after ESD or RFA (Barrett's esophagus) were treated [2]. The favorable outcomes of these studies will be published soon (manuscripts in preparation) and a randomized clinical trial is planned. These clinical studies, by virtue of in vitro experiments demonstrating the activation of stem cells and using confocal laser microscopy to monitor the healing of these cell sheets, underscore the feasibility and rationale of such an approach [3]. I declare that Masayuki Yamato is a shareholder of CellSeed Inc. References
- Published
- 2015
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