41 results on '"Invernici, G"'
Search Results
2. Dermal fibroblasts display similar phenotypic and differentiation capacity to fat-derived mesenchymal stem cells, but differ in anti-inflammatory and angiogenic potential: OP-008
- Author
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Blasi, A, Balducci, L, Soleti, A, Invernici, G, Parati, E, and Alessandri, G
- Published
- 2011
3. Anthropometric measurements, handgrip strength and bioelectrical impedance analysis in assessment of the nutritional status of ascitic patients with nonalcoholic liver cirrhosis awaiting liver transplantation
- Author
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CORTINOVIS, F., CORTESI, L., BRESCIANI, L., INVERNICI, G., VERGA, G., GAFFURI, G., LUCÀ, M. G., PASULO, L., COLPANI, M., FAGIUOLI, S., and SILEO, F.
- Published
- 2009
4. Human microvascular endothelial cells from different fetal organs demonstrate organ-specific CAM expression
- Author
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Invernici, G., Ponti, D., Corsini, E., Cristini, S., Frigerio, S., Colombo, A., Parati, E., and Alessandri, G.
- Published
- 2005
- Full Text
- View/download PDF
5. Engraftment, neuroglial transdifferentiation and behavioral recovery after complete spinal cord transection in rats
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Giacomo Rossi, Luca Lacitignola, Antonio Crovace, E. Francioso, Sabino Luzzi, Alberto Maria Crovace, Valentini, Renato Galzio, and Invernici G
- Subjects
0301 basic medicine ,red fluorescent protein ,Pathology ,medicine.medical_specialty ,Neurofilament ,03 medical and health sciences ,Head and Spinal Cord Transplantation: Original Article ,0302 clinical medicine ,medicine ,Spinal cord injury ,neurological recovery ,neuroglial differentiation ,business.industry ,Mesenchymal stem cell ,Transdifferentiation ,Nestin ,medicine.disease ,Spinal cord ,Neuroregeneration ,spinal cord injury ,030104 developmental biology ,medicine.anatomical_structure ,Mesenchymal stem cells ,Surgery ,Neurology (clinical) ,Bone marrow ,Erratum ,business ,030217 neurology & neurosurgery - Abstract
Background: Proof of the efficacy and safety of a xenogeneic mesenchymal stem cell (MSCs) transplant for spinal cord injury (SCI) may theoretically widen the spectrum of possible grafts for neuroregeneration. Methods: Twenty rats were submitted to complete spinal cord transection. Ovine bone marrow MSCs, retrovirally transfected with red fluorescent protein and not previously induced for neuroglial differentiation, were applied in 10 study rats (MSCG). Fibrin glue was injected in 10 control rats (FGG). All rats were evaluated on a weekly basis and scored using the Basso–Beattie–Bresnahan (BBB) locomotor scale for 10 weeks, when the collected data were statistically analyzed. The spinal cords were then harvested and analyzed with light microscopy, immunohistochemistry, and immunofluorescence. Results: Ovine MSCs culture showed positivity for Nestin. MSCG had a significant and durable recovery of motor functions (P
- Published
- 2018
6. Anthropopmetric measurements, hand grip, strenght and bioelectrical impedance analysis in assessment of the nutritional status of ascitic patients with non-alcoholic liver cirrhosis awaiting for liver transplantation
- Author
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Cortinovis, F, Cortesi, L, Brescianil, Invernici, G, Verga, G, Gaffuri, G, Lucà, M, Pasulo, L, Colpani, M, Fagiuoli, S, Sileo, F, Cortinovis F, Cortesi L, BrescianiL, Invernici G, Verga G, Gaffuri G, Lucà MG, Pasulo L, Colpani M, Fagiuoli S, Sileo F, Cortinovis, F, Cortesi, L, Brescianil, Invernici, G, Verga, G, Gaffuri, G, Lucà, M, Pasulo, L, Colpani, M, Fagiuoli, S, Sileo, F, Cortinovis F, Cortesi L, BrescianiL, Invernici G, Verga G, Gaffuri G, Lucà MG, Pasulo L, Colpani M, Fagiuoli S, and Sileo F
- Abstract
Background: Malnutrition is highly prevalent among patients awaiting liver transplantation. Patients with poor nutritional status before transplant will have more complications and higher mortality postoperatively. Nutritional assessment should be performed in every patient with chronic liver disease awaiting liver transplantation in order to ensure correct and timely nutritional treatment, punctual monitoring pre- and post-transplant, and adequate global prognostic evaluation of the patient. Objective: The aim of the study was to identify a reliable and sensitive panel of measurements for the assessment of malnutrition in patients with chronic liver disease, which could also predict postoperative complications in orthotopic liver transplant recipients.Patients and methods: Twenty-five consecutive hospitalized ascitic patients (18 men, 7 women) with nonalcoholic liver cirrhosis were assessed with anthropometric measurements, handgrip strength and bioelectrical impedance analysis. Result: Comparison between the observed values (measured and calculated) in our 25 patients and the expected values based on percentile distribution showed statistically significant differences for the parameters triceps skinfold thickness, midarm circumference, arm muscle area, fat-free mass, handgrip strength and phase angle. Conclusion: The nutritional evaluation of cirrhotic patients awaiting liver transplant must be as exhaustive as possible. Bioelectrical impedance analysis adds a further parameter to anthropomet- ric measurements and handgrip strength, namely the phase angle, which provides usefulprognostic indications for the selection of candidates for liver transplantation.
- Published
- 2009
7. Anthropopmetric measurements, hand grip, strenght and bioelectrical impedance analysis in assessment of the nutritional status of ascitic patients with non-alcoholic liver cirrhosis awaiting for liver transplantation
- Author
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Cortinovis F, Cortesi L, BrescianiL, Invernici G, Verga G, Gaffuri G, Lucà MG, Pasulo L, Colpani M, Fagiuoli S, Sileo F, Cortinovis, F, Cortesi, L, Brescianil, Invernici, G, Verga, G, Gaffuri, G, Lucà, M, Pasulo, L, Colpani, M, Fagiuoli, S, and Sileo, F
- Subjects
Anthropometric assessment ,Phase angle ,Liver transplantation ,Bioelectrical impedance analysi ,Nutritional statu ,Handgrip strength ,Liver cirrhosi - Abstract
Background: Malnutrition is highly prevalent among patients awaiting liver transplantation. Patients with poor nutritional status before transplant will have more complications and higher mortality postoperatively. Nutritional assessment should be performed in every patient with chronic liver disease awaiting liver transplantation in order to ensure correct and timely nutritional treatment, punctual monitoring pre- and post-transplant, and adequate global prognostic evaluation of the patient. Objective: The aim of the study was to identify a reliable and sensitive panel of measurements for the assessment of malnutrition in patients with chronic liver disease, which could also predict postoperative complications in orthotopic liver transplant recipients.Patients and methods: Twenty-five consecutive hospitalized ascitic patients (18 men, 7 women) with nonalcoholic liver cirrhosis were assessed with anthropometric measurements, handgrip strength and bioelectrical impedance analysis. Result: Comparison between the observed values (measured and calculated) in our 25 patients and the expected values based on percentile distribution showed statistically significant differences for the parameters triceps skinfold thickness, midarm circumference, arm muscle area, fat-free mass, handgrip strength and phase angle. Conclusion: The nutritional evaluation of cirrhotic patients awaiting liver transplant must be as exhaustive as possible. Bioelectrical impedance analysis adds a further parameter to anthropomet- ric measurements and handgrip strength, namely the phase angle, which provides usefulprognostic indications for the selection of candidates for liver transplantation.
- Published
- 2009
8. Transforming growth factor-beta1 and CD105 promote the migration of hepatocellular carcinoma-derived endothelium
- Author
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Benetti, Anna, Berenzi, Angiola, Gambarotti, M, Garrafa, Emirena Michela, Gelati, M, Dessy, Enrico, Portolani, Nazario, Piardi, Tullio, Giulini, Stefano Maria, Caruso, Arnaldo, Invernici, G, Parati, E. A., Nicosia, R, and Alessandri, G.
- Published
- 2008
9. HUMAN ADULT SKELETAL MUSCLE STEM CELLS DIFFERENTIATE INTOCARDIOMYOCYTE AND IMPROVE LV FUNCTION IN AN ISCHEMICINJURY MODEL
- Author
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Piccoli, P., Bisleri, Gianluigi, Invernici, G., Madeddu, P., Fascio, U., Alessandri, G., Parati, E., and Muneretto, Claudio
- Published
- 2008
10. Melanoma contains CD133 and ABCG2 positive cells with enhanced tumourigenic potential
- Author
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Monzani, E, Facchetti, F, Galmozzi, E, Corsini, E, Benetti, A, Cavazzin, C, Gritti, A, Piccinini, A, Porro, D, Santinami, M, Invernici, G, Parati, E, Alessandri, G, La Porta, C, La Porta, CA, PORRO, DANILO, Monzani, E, Facchetti, F, Galmozzi, E, Corsini, E, Benetti, A, Cavazzin, C, Gritti, A, Piccinini, A, Porro, D, Santinami, M, Invernici, G, Parati, E, Alessandri, G, La Porta, C, La Porta, CA, and PORRO, DANILO
- Abstract
The failure to eradicate most cancers and in particular melanoma may be as fundamental as a misidentification of the target. The identification of cancer stem/initiating cells within the tumour population with a crucial role for tumour formation may open new pharmacological perspectives. Our data show three main novelties for human melanoma: firstly, melanoma biopsy contains a subset of cells expressing CD133 (CD133+) and the latter is able to develop a Mart-1 positive tumour in NOD-SCID mice. Secondly, the WM115, a human melanoma cell line, has been found to express both CD133 and ABCG2 markers. This cell line grows as floating spheroids, expresses typical progenitors and mature neuronal/oligodendrocyte markers and is able to transdifferentiate into astrocytes or mesenchymal lineages under specific growth conditions. As in xenografts generated with CD133+ biopsy melanoma cells, those produced by the cell line displayed lower levels of CD133 and ABCG2. Thirdly, the WM115 cells express the most important angiogenic and lymphoangiogenic factors such as notch 4, prox1 and podoplanin which can cooperate in the development of the tumourigenic capability of melanoma in vivo. Therefore, in this study, we demonstrate the presence of stem/initiating subsets in melanoma both in biopsy and in an established melanoma cell line grown in vitro and in xenografts. Interestingly, considering that melanoma gives metastasis primarily through lymphatic vessels, herein, we demonstrated that a melanoma cell line expresses typical lymphoangiogenic factors.
- Published
- 2007
11. HUMAN SKELETAL MUSCLE-DERIVED STEM CELLS FOR MYOCAR-DIAL REGENERATION
- Author
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Bisleri, C, primary, Alessandri, C, additional, Invernici, G, additional, Negri, A, additional, Manfredi, J, additional, Caruso, A, additional, and Muneretto, C, additional
- Published
- 2004
- Full Text
- View/download PDF
12. An innovative treatment for cardiac failure: Autologous stem cell transplantation
- Author
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Invernici, G., Cristini, S., Navone, S., Canzi, L., and Eugenio Agostino Parati
13. Valutazione antropometrica, dinamometrica ed impedenziometrica dello stato nutrizionale nel paziente con cirrosi postepatitica, ascitico, candidato a trapianto di fegato.
- Author
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Cortinovis F, Cortesi L, Bresciani L, Invernici G, Verga G, Gaffuri G, Luca MG, Pasulo L, Colpani M, Fagiuoli S, and Sileo F
- Published
- 2009
14. Tumour vascularization via endothelial differentiation of glioblastoma stem-like cells
- Author
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Roberto Pallini, Mauro Biffoni, Eugenio Parati, Giorgio Stassi, Ruggero De Maria, Giulio Maira, Luigi Maria Larocca, Matilde Todaro, Tonia Cenci, Lucia Ricci-Vitiani, Gloria Invernici, Ricci-Vitiani, L, Pallini, R, Biffoni, M, Todaro, M, Invernici, G, Cenci, T, Maira, G, Parati, EA, Stassi, G, Larocca, LM, and De Maria, R
- Subjects
Endothelium ,Angiogenesis ,Transplantation, Heterologous ,Settore MED/27 - NEUROCHIRURGIA ,Mice, Transgenic ,Mice, SCID ,Biology ,Models, Biological ,Mice ,Vasculogenesis ,Neural Stem Cells ,Mice, Inbred NOD ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Cell Lineage ,Vasculogenic mimicry ,glioblastoma, tumor vascularization ,In Situ Hybridization, Fluorescence ,Chromosome Aberrations ,Multidisciplinary ,Neovascularization, Pathologic ,Endothelial Cells ,Cell Differentiation ,Vascular endothelial growth factor B ,Endothelial stem cell ,Vascular endothelial growth factor A ,medicine.anatomical_structure ,Vascular endothelial growth factor C ,Tumor Markers, Biological ,Immunology ,Cancer research ,Endothelium, Vascular ,Glioblastoma ,Neoplasm Transplantation - Abstract
Glioblastoma is a highly angiogenetic malignancy, the neoformed vessels of which are thought to arise by sprouting of pre-existing brain capillaries. The recent demonstration that a population of glioblastoma stem-like cells (GSCs) maintains glioblastomas indicates that the progeny of these cells may not be confined to the neural lineage. Normal neural stem cells are able to differentiate into functional endothelial cells. The connection between neural stem cells and the endothelial compartment seems to be critical in glioblastoma, where cancer stem cells closely interact with the vascular niche and promote angiogenesis through the release of vascular endothelial growth factor(VEGF) and stromal-derived factor 1 (refs 5-9). Here we show that a variable number (range 20-90%, mean 60.7%) of endothelial cells in glioblastoma carry the same genomic alteration as tumour cells, indicating that a significant portion of the vascular endothelium has a neoplastic origin. The vascular endothelium contained a subset of tumorigenic cells that produced highly vascularized anaplastic tumours with areas of vasculogenic mimicry in immunocompromised mice. In vitro culture of GSCs in endothelial conditions generated progeny with phenotypic and functional features of endothelial cells. Likewise, orthotopic or subcutaneous injection of GSCs in immunocompromised mice produced tumour xenografts, the vessels of which were primarily composed of human endothelial cells. Selective targeting of endothelial cells generated by GSCs in mouse xenografts resulted in tumour reduction and degeneration, indicating the functional relevance of the GSC-derived endothelial vessels. These findings describe a new mechanism for tumour vasculogenesis and may explain the presence of cancer-derived endothelial-like cells in several malignancies.
- Published
- 2010
15. Human Fetal Aorta Contains Vascular Progenitor Cells Capable of Inducing Vasculogenesis, Angiogenesis, and Myogenesis in Vitro and in a Murine Model of Peripheral Ischemia
- Author
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Cesare Peschle, Gloria Invernici, Tommaso Rizzuti, Giulio Alessandri, Umberto Fascio, Eugenio Parati, Mauro Siragusa, Costanza Emanueli, Silvia Cristini, Paolo Madeddu, Anna Benetti, Roberto F. Nicosia, Sergio Domenico Gadau, Emilio Ciusani, Giorgio Stassi, Augusto Colombo, INVERNICI G, EMANUELI C, MADEDDU P, CRISTINI S, GADAU S, BENETTI A, CIUSANI E, STASSI G, SIRAGUSA M, NICOSIA R, PESCHLE C, FASCIO U, COLOMBO A, RIZZUTI T, PARATI E, and ALESSANDRI G
- Subjects
Vascular Endothelial Growth Factor A ,Angiogenesis ,Becaplermin ,Neovascularization, Physiologic ,Antigens, CD34 ,Biology ,Muscle Development ,Mural cell ,Pathology and Forensic Medicine ,Angiopoietin-2 ,Mice ,Fetus ,Vasculogenesis ,Antigens, CD ,Ischemia ,Animals ,Humans ,Cell Lineage ,AC133 Antigen ,Progenitor cell ,Aorta ,Cells, Cultured ,Glycoproteins ,Platelet-Derived Growth Factor ,Stem Cells ,Proto-Oncogene Proteins c-sis ,Vascular Endothelial Growth Factor Receptor-2 ,Cell biology ,Endothelial stem cell ,Vascular endothelial growth factor B ,Vascular endothelial growth factor A ,Vascular endothelial growth factor C ,Immunology ,Blood Vessels ,Peptides ,Biomarkers ,Regular Articles - Abstract
Vasculogenesis, the formation of blood vessels in embryonic or fetal tissue mediated by immature vascular cells (ie, angioblasts), is poorly understood. We report the identification of a population of vascular progenitor cells (hVPCs) in the human fetal aorta composed of undifferentiated mesenchymal cells that coexpress endothelial and myogenic markers. Under culture conditions that promoted cell differentiation, hVPCs gave rise to a mixed population of mature endothelial and mural cells when progenitor cells were stimulated with vascular endothelial growth factor-A or platelet-derived growth factor-betabeta. hVPCs grew as nonadherent cells and, when embedded in a three-dimensional collagen gel, reorganized into cohesive cellular cords that resembled mature vascular structures. hVPC-conditioned medium contained angiogenic substances (vascular endothelial growth factor-A and angiopoietin-2) and strongly stimulated the proliferation of endothelial cells. We also demonstrate the therapeutic efficacy of a small number of hVPCs transplanted into ischemic limb muscle of immunodeficient mice. hVPCs markedly improved neovascularization and inhibited the loss of endogenous endothelial cells and myocytes, thus ameliorating the clinical outcome from ischemia. We conclude that fetal aorta represents an important source for the investigation of the phenotypic and functional features of human vascular progenitor cells.
- Published
- 2007
16. Melanoma contains CD133 and ABCG2 positive cells with enhanced tumourigenic potential
- Author
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Gloria Invernici, Chiara Cavazzin, Andrea Piccinini, Giulio Alessandri, Anna Benetti, mario santinami, Floriana Facchetti, Enrico Galmozzi, Angela Gritti, Caterina A. M. La Porta, Danilo Porro, Eugenio Parati, Elena Monzani, Elena Corsini, Monzani, E, Facchetti, F, Galmozzi, E, Corsini, E, Benetti, A, Cavazzin, C, Gritti, A, Piccinini, A, Porro, D, Santinami, M, Invernici, G, Parati, E, Alessandri, G, and La Porta, C
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Skin Neoplasms ,Blotting, Western ,Transplantation, Heterologous ,Population ,Mice, SCID ,Biology ,Metastasis ,Mice ,Antigens, CD ,Mice, Inbred NOD ,Cancer stem cell ,Tumor Cells, Cultured ,medicine ,ATP Binding Cassette Transporter, Subfamily G, Member 2 ,Animals ,AC133 Antigen ,CD133 ,Progenitor cell ,education ,Melanoma ,neoplasms ,Glycoproteins ,education.field_of_study ,Neovascularization, Pathologic ,Reverse Transcriptase Polymerase Chain Reaction ,Cancer stem cells ,Mesenchymal stem cell ,ABCB5 ,medicine.disease ,Immunohistochemistry ,Neoplasm Proteins ,Oncology ,embryonic structures ,Cancer research ,ATP-Binding Cassette Transporters ,Stem cell ,Peptides ,Biomarkers - Abstract
The failure to eradicate most cancers and in particular melanoma may be as fundamental as a misidentification of the target. The identification of cancer stem/initiating cells within the tumour population with a crucial role for tumour formation may open new pharmacological perspectives. Our data show three main novelties for human melanoma: firstly, melanoma biopsy contains a subset of cells expressing CD133 (CD133+) and the latter is able to develop a Mart-1 positive tumour in NOD-SCID mice. Secondly, the WM115, a human melanoma cell line, has been found to express both CD133 and ABCG2 markers. This cell line grows as floating spheroids, expresses typical progenitors and mature neuronal/oligodendrocyte markers and is able to transdifferentiate into astrocytes or mesenchymal lineages under specific growth conditions. As in xenografts generated with CD133+ biopsy melanoma cells, those produced by the cell line displayed lower levels of CD133 and ABCG2. Thirdly, the WM115 cells express the most important angiogenic and lymphoangiogenic factors such as notch 4, prox1 and podoplanin which can cooperate in the development of the tumourigenic capability of melanoma in vivo. Therefore, in this study, we demonstrate the presence of stem/initiating subsets in melanoma both in biopsy and in an established melanoma cell line grown in vitro and in xenografts. Interestingly, considering that melanoma gives metastasis primarily through lymphatic vessels, herein, we demonstrated that a melanoma cell line expresses typical lymphoangiogenic factors.
- Published
- 2007
17. Human and mouse brain-derived endothelial cells require high levels of growth factors medium for their isolation, in vitro maintenance and survival
- Author
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Eugenio Parati, Giulio Alessandri, Silvia Cristini, Stefania Elena Navone, S. Sangiorgi, Gloria Invernici, Emilio Ciusani, Sara Nava, Sergio Balbi, Alessandra Bosutti, Mark Slevin, Giovanni Marfia, Navone, Se, Marfia, G, Nava, S, Invernici, G, Cristini, S, Balbi, S, Sangiorgi, S, Ciusani, E, Bosutti, Alessandra, Alessandri, G, Slevin, M, and Parati, E. A.
- Subjects
CD31 ,Pathology ,medicine.medical_specialty ,Proliferation index ,Computer Networks and Communications ,Brain microvascular endothelial cell ,Blood–brain barrier ,Brain microvascular endothelial cells ,Developmental Neuroscience ,In vivo ,Endothelial permeability ,medicine ,ITGAV ,Endothelial junction ,business.industry ,Research ,Endothelial junctions ,Blood brain barrier ,Cell Biology ,Endoglin ,In vitro ,Cell biology ,medicine.anatomical_structure ,Neurology ,CD146 ,business - Abstract
BACKGROUND: Brain microvascular endothelial cells (BMVECs) constitute the primary limitation for passage of ions and molecules from the blood into the brain through the blood brain barrier. Numerous multi-step procedures for isolating and culturing BMVECs have been described. However, each one demonstrates major limitations in purity of culture and/or low proliferation rate. Our goal was to study the efficiency of our pending patent medium, Endothelial Proliferation Medium (EndoPM), on the isolation and purification of human and murine BMVECs. METHODS: BMVECs, cultured in EndoPM were compared to those cultured in a commercial medium EBM. Cultures were characterized by flow cytometric analysis, lineage differentiation, the ability to form tube-like structure, immunofluorescence, molecular analyses and also in an in vivo model assay. Moreover permeability was assayed by monitoring the passage of Dextran-FITC through a tight monolayer of BMVECs grown to confluence in Boyden chambers. One way Anova two-tailed test was utilized for all statistical analyses. RESULTS: The properties of ECs in human and murine BMVECs is confirmed by the expression of endothelial markers (CD31, CD105, CD146, Tie-2 and vWF), of representative proangiogenic genes (ICAM1, VCAM1 and integrin ITGAV), of considerable tube-forming ability, with low-density lipoprotein uptake, eNOS and GLUT-1 expression. Furthermore cells are able to express markers of the junctional architecture as VE-cadherin, β-catenin and Claudin-5 and greatly reduce dextran permeability as barrier functional test. Moreover BMVECs spontaneously organize in vascular-like structures and maintain the expression of endothelial markers in an in vivo xenograft model assay. The significant effect of EndoPM is confirmed by the study of proliferation index, survival index and the behaviour of BMVECs and fibroblasts in co-culture conditions. CONCLUSION: Herein we describe a simple and reproducible method for the isolation and expansion of human and mouse BMVECs, based on a newly formulated medium (EndoPM) with optimized concentration of growth factors (EGF, FGF-2 and Bovine Brain Extract-BBE). This procedure should facilitate the isolation and expansion of human and mouse BMVECs with extended lifetime, good viability and purity. This approach may provide an effective strategy to aid phenotypical and functional studies of brain vessels under physiological and pathological conditions.
- Published
- 2013
18. Benfotiamine accelerates the healing of ischaemic diabetic limbs in mice through protein kinase B/Akt-mediated potentiation of angiogenesis and inhibition of apoptosis
- Author
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Ilaria Campesi, Gallia Graiani, Costanza Emanueli, K. Ward, S Van Linthout, Frank Spillmann, Matilde Todaro, Paolo Madeddu, Gloria Invernici, Sergio Domenico Gadau, Marco Meloni, Gadau, S, Emanueli, C, Van Linthout, S, Graiani, G, Todaro, M, Meloni, M, Campesi, I, Invernici, G, Spillmann, F, Ward, K, and Madeddu, P
- Subjects
Male ,medicine.medical_specialty ,Programmed cell death ,Angiogenesis ,Endocrinology, Diabetes and Metabolism ,Blotting, Western ,Neovascularization, Physiologic ,Apoptosis ,Mice, Inbred Strains ,Biology ,Diabetes Mellitus, Experimental ,Neovascularization ,Mice ,Random Allocation ,Ischemia ,Internal medicine ,Internal Medicine ,medicine ,Animals ,Thiamine ,Muscle, Skeletal ,Protein kinase B ,Cell Proliferation ,Caspase 3 ,Stem Cells ,protein kinase, PKB/Akt ,Body Weight ,Hemodynamics ,Endothelial Cells ,Caspase Inhibitors ,Immunohistochemistry ,Endothelial stem cell ,Enzyme Activation ,Oxidative Stress ,Endocrinology ,Benfotiamine ,Caspases ,Dietary Supplements ,Transketolase activity ,medicine.symptom ,Proto-Oncogene Proteins c-akt ,Diabetic Angiopathies ,medicine.drug - Abstract
Benfotiamine, a vitamin B1 analogue, reportedly prevents diabetic microangiopathy. The aim of this study was to evaluate whether benfotiamine is of benefit in reparative neovascularisation using a type I diabetes model of hindlimb ischaemia. We also investigated the involvement of protein kinase B (PKB)/Akt in the therapeutic effects of benfotiamine. Streptozotocin-induced diabetic mice, given oral benfotiamine or vehicle, were subjected to unilateral limb ischaemia. Reparative neovascularisation was analysed by histology. The expression of Nos3 and Casp3 was evaluated by real-time PCR, and the activation state of PKB/Akt was assessed by western blot analysis and immunohistochemistry. The functional importance of PKB/Akt in benfotiamine-induced effects was investigated using a dominant-negative construct. Diabetic muscles showed reduced transketolase activity, which was corrected by benfotiamine. Importantly, benfotiamine prevented ischaemia-induced toe necrosis, improved hindlimb perfusion and oxygenation, and restored endothelium-dependent vasodilation. Histological studies revealed the improvement of reparative neovascularisation and the inhibition of endothelial and skeletal muscle cell apoptosis. In addition, benfotiamine prevented the vascular accumulation of advanced glycation end products and the induction of pro-apoptotic caspase-3, while restoring proper expression of Nos3 and Akt in ischaemic muscles. The benefits of benfotiamine were nullified by dominant-negative PKB/Akt. In vitro, benfotiamine stimulated the proliferation of human EPCs, while inhibiting apoptosis induced by high glucose. In diabetic mice, the number of circulating EPCs was reduced, with the deficit being corrected by benfotiamine. We have demonstrated, for the first time, that benfotiamine aids the post-ischaemic healing of diabetic animals via PKB/Akt-mediated potentiation of angiogenesis and inhibition of apoptosis. In addition, benfotiamine combats the diabetes-induced deficit in endothelial progenitor cells.
- Published
- 2005
19. Fresh lemon juice supplementation for the prevention of recurrent stones in calcium oxalate nephrolithiasis: A pragmatic, prospective, randomised, open, blinded endpoint (PROBE) trial.
- Author
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Ruggenenti P, Caruso MR, Cortinovis M, Perna A, Peracchi T, Giuliano GA, Rota S, Brambilla P, Invernici G, Villa D, Diadei O, Trillini M, Natali G, and Remuzzi G
- Abstract
Background: Standard diet with normal calcium and reduced animal proteins and salt content reduces stone recurrence in calcium oxalate nephrolithiasis. Whether lemon juice supplementation further reduces recurrence rate is unknown., Methods: In this single-centre, prospective, randomised, open, blinded endpoint trial (Clinical Trials gov NCT01217372) we evaluated the effects of fresh lemon juice supplementation (60 mL twice daily) versus no supplementation, on time to stone recurrence in 203 patients with recurrent idiopathic calcium oxalate nephrolithiasis who were all prescribed a standard diet. Patients were included between July 2009 and March 2017 at the Nephrology Unit of the Papa Giovanni XXIII hospital in Bergamo, Italy. Time to stone recurrence at 2 years of follow-up was the primary outcome. Analyses were by intention-to-treat., Findings: During two years of follow-up 21 of 100 patients randomised to lemon juice supplementation and 32 of 103 controls randomised to no supplementation had stone recurrence [HR (95% CI): 0·62 (0·35-1·07), p = 0·089]. Patient adherence to lemon juice supplementation, however, progressively decreased from 68% at one-year to 48% at two-year follow-up. At explorative analyses restricted at one-year follow-up, ten patients with supplementation versus 22 controls had stone recurrence [0·43 (0·20-0·89), p = 0·028]. After adjustment by age, sex and normo or hypocitraturia, the HR (95%) was still significant [0·45 (0·20-0·93), p = 0·036]. At six months, 24 hour urinary sodium excretion decreased by 8·60±65·68 mEq/24 h in patients receiving lemon juice supplementation and increased by 3·88±64·78 mEq/24 h in controls. Changes significantly differed between groups ( p = 0·031). This difference was subsequently lost. Treatment was safe. In patients with lemon juice supplementation gastrointestinal disorders were more frequent ( p <0·001). Renal and urinary tract disorders were similar between groups ( p = 0·103)., Interpretation: Explorative analyses suggest that f resh lemon juice supplementation to standard diet might prevent stone recurrence in patients with calcium-oxalate nephrolithiasis. However, treatment effect was likely reduced by progressively declining adherence to lemon juice supplementation., Funding: This study received no funding., Competing Interests: G.R. reported personal fees from Akebia Pharmaceuticals Inc, Alexion Pharmaceuticals, BioCryst Pharmaceuticals Ins, AstraZeneca and Janssen Research & Development LLC, as well as speaker honorarium/travel reimbursements from Boehringer Ingelheim, Menarini Ricerche Spa and Silence Therapeutics. All the other authors declare that they have nothing to disclose., (© 2021 The Authors.)
- Published
- 2021
- Full Text
- View/download PDF
20. Engraftment, neuroglial transdifferentiation and behavioral recovery after complete spinal cord transection in rats.
- Author
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Luzzi S, Crovace AM, Lacitignola L, Valentini V, Francioso E, Rossi G, Invernici G, Galzio RJ, and Crovace A
- Abstract
Background: Proof of the efficacy and safety of a xenogeneic mesenchymal stem cell (MSCs) transplant for spinal cord injury (SCI) may theoretically widen the spectrum of possible grafts for neuroregeneration., Methods: Twenty rats were submitted to complete spinal cord transection. Ovine bone marrow MSCs, retrovirally transfected with red fluorescent protein and not previously induced for neuroglial differentiation, were applied in 10 study rats (MSCG). Fibrin glue was injected in 10 control rats (FGG). All rats were evaluated on a weekly basis and scored using the Basso-Beattie-Bresnahan (BBB) locomotor scale for 10 weeks, when the collected data were statistically analyzed. The spinal cords were then harvested and analyzed with light microscopy, immunohistochemistry, and immunofluorescence., Results: Ovine MSCs culture showed positivity for Nestin. MSCG had a significant and durable recovery of motor functions ( P <.001). Red fluorescence was found at the injury sites in MSCG. Positivity for Nestin, tubulin βIII, NG2 glia, neuron-specific enolase, vimentin, and 200 kD neurofilament were also found at the same sites., Conclusions: Xenogeneic ovine bone marrow MSCs proved capable of engrafting into the injured rat spinal cord. Transdifferentiation into a neuroglial phenotype was able to support partial functional recovery., Competing Interests: There are no conflicts of interest.
- Published
- 2018
- Full Text
- View/download PDF
21. Decellularized silk fibroin scaffold primed with adipose mesenchymal stromal cells improves wound healing in diabetic mice.
- Author
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Navone SE, Pascucci L, Dossena M, Ferri A, Invernici G, Acerbi F, Cristini S, Bedini G, Tosetti V, Ceserani V, Bonomi A, Pessina A, Freddi G, Alessandrino A, Ceccarelli P, Campanella R, Marfia G, Alessandri G, and Parati EA
- Subjects
- Adipose Tissue cytology, Animals, Cell Adhesion, Cell Movement, Cell Proliferation, Cells, Cultured, Fibroblasts physiology, Human Umbilical Vein Endothelial Cells physiology, Humans, Keratinocytes physiology, Mesenchymal Stem Cells drug effects, Mesenchymal Stem Cells physiology, Mice, Mice, Obese, Neovascularization, Physiologic, Rats, Rats, Sprague-Dawley, Receptors, Leptin genetics, Fibroins pharmacology, Mesenchymal Stem Cell Transplantation methods, Mesenchymal Stem Cells cytology, Re-Epithelialization, Tissue Scaffolds chemistry
- Abstract
Introduction: Silk fibroin (SF) scaffolds have been shown to be a suitable substrate for tissue engineering and to improve tissue regeneration when cellularized with mesenchymal stromal cells (MSCs). We here demonstrate, for the first time, that electrospun nanofibrous SF patches cellularized with human adipose-derived MSCs (Ad-MSCs-SF), or decellularized (D-Ad-MSCs-SF), are effective in the treatment of skin wounds, improving skin regeneration in db/db diabetic mice., Methods: The conformational and structural analyses of SF and D-Ad-MSCs-SF patches were performed by scanning electron microscopy, confocal microscopy, Fourier transform infrared spectroscopy and differential scanning calorimetry. Wounds were performed by a 5 mm punch biopsy tool on the mouse's back. Ad-MSCs-SF and D-Ad-MSCs-SF patches were transplanted and the efficacy of treatments was assessed by measuring the wound closure area, by histological examination and by gene expression profile. We further investigated the in vitro angiogenic properties of Ad-MSCs-SF and D-Ad-MSCs-SF patches by affecting migration of human umbilical vein endothelial cells (HUVECs), keratinocytes (KCs) and dermal fibroblasts (DFs), through the aortic ring assay and, finally, by evaluating the release of angiogenic factors., Results: We found that Ad-MSCs adhere and grow on SF, maintaining their phenotypic mesenchymal profile and differentiation capacity. Conformational and structural analyses on SF and D-Ad-MSCs-SF samples, showed that sterilization, decellularization, freezing and storing did not affect the SF structure. When grafted in wounds of diabetic mice, both Ad-MSCs-SF and D-Ad-MSCs-SF significantly improved tissue regeneration, reducing the wound area respectively by 40% and 35%, within three days, completing the process in around 10 days compared to 15-17 days of controls. RT2 gene profile analysis of the wounds treated with Ad-MSCs-SF and D-Ad-MSCs-SF showed an increment of genes involved in angiogenesis and matrix remodeling. Finally, Ad-MSCs-SF and D-Ad-MSCs-SF co-cultured with HUVECs, DFs and KCs, preferentially enhanced the HUVECs' migration and the release of angiogenic factors stimulating microvessel outgrowth in the aortic ring assay., Conclusions: Our results highlight for the first time that D-Ad-MSCs-SF patches are almost as effective as Ad-MSCs-SF patches in the treatment of diabetic wounds, acting through a complex mechanism that involves stimulation of angiogenesis. Our data suggest a potential use of D-Ad-MSCs-SF patches in chronic diabetic ulcers in humans.
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- 2014
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22. Isolation and expansion of human and mouse brain microvascular endothelial cells.
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Navone SE, Marfia G, Invernici G, Cristini S, Nava S, Balbi S, Sangiorgi S, Ciusani E, Bosutti A, Alessandri G, Slevin M, and Parati EA
- Subjects
- Animals, Cell Survival, Coculture Techniques, Flow Cytometry, Fluorescent Antibody Technique, Humans, Mice, Neovascularization, Physiologic, Cell Culture Techniques, Endothelial Cells cytology, Microvessels cytology
- Abstract
Brain microvascular endothelial cells (BMVECs) have an important role in the constitution of the blood-brain barrier (BBB). The BBB is involved in the disease processes of a number of neurological disorders in which its permeability increases. Isolation of BMVECs could elucidate the mechanism involved in these processes. This protocol describes how to isolate and expand human and mouse BMVECs. The procedure covers brain-tissue dissociation, digestion and cell selection. Cells are selected on the basis of time-responsive differential adhesiveness to a collagen type I-precoated surface. The protocol also describes immunophenotypic characterization, cord formation and functional assays to confirm that these cells in endothelial proliferation medium (EndoPM) have an endothelial origin. The entire technique requires ∼7 h of active time. Endothelial cell clusters are readily visible after 48 h, and expansion of BMVECs occurs over the course of ∼60 d.
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- 2013
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23. Human and mouse brain-derived endothelial cells require high levels of growth factors medium for their isolation, in vitro maintenance and survival.
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Navone SE, Marfia G, Nava S, Invernici G, Cristini S, Balbi S, Sangiorgi S, Ciusani E, Bosutti A, Alessandri G, Slevin M, and Parati EA
- Abstract
Background: Brain microvascular endothelial cells (BMVECs) constitute the primary limitation for passage of ions and molecules from the blood into the brain through the blood brain barrier. Numerous multi-step procedures for isolating and culturing BMVECs have been described. However, each one demonstrates major limitations in purity of culture and/or low proliferation rate. Our goal was to study the efficiency of our pending patent medium, Endothelial Proliferation Medium (EndoPM), on the isolation and purification of human and murine BMVECs., Methods: BMVECs, cultured in EndoPM were compared to those cultured in a commercial medium EBM. Cultures were characterized by flow cytometric analysis, lineage differentiation, the ability to form tube-like structure, immunofluorescence, molecular analyses and also in an in vivo model assay. Moreover permeability was assayed by monitoring the passage of Dextran-FITC through a tight monolayer of BMVECs grown to confluence in Boyden chambers. One way Anova two-tailed test was utilized for all statistical analyses., Results: The properties of ECs in human and murine BMVECs is confirmed by the expression of endothelial markers (CD31, CD105, CD146, Tie-2 and vWF), of representative proangiogenic genes (ICAM1, VCAM1 and integrin ITGAV), of considerable tube-forming ability, with low-density lipoprotein uptake, eNOS and GLUT-1 expression. Furthermore cells are able to express markers of the junctional architecture as VE-cadherin, β-catenin and Claudin-5 and greatly reduce dextran permeability as barrier functional test. Moreover BMVECs spontaneously organize in vascular-like structures and maintain the expression of endothelial markers in an in vivo xenograft model assay. The significant effect of EndoPM is confirmed by the study of proliferation index, survival index and the behaviour of BMVECs and fibroblasts in co-culture conditions., Conclusion: Herein we describe a simple and reproducible method for the isolation and expansion of human and mouse BMVECs, based on a newly formulated medium (EndoPM) with optimized concentration of growth factors (EGF, FGF-2 and Bovine Brain Extract-BBE). This procedure should facilitate the isolation and expansion of human and mouse BMVECs with extended lifetime, good viability and purity. This approach may provide an effective strategy to aid phenotypical and functional studies of brain vessels under physiological and pathological conditions.
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- 2013
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24. Contribution of novel ATGL missense mutations to the clinical phenotype of NLSD-M: a strikingly low amount of lipase activity may preserve cardiac function.
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Tavian D, Missaglia S, Redaelli C, Pennisi EM, Invernici G, Wessalowski R, Maiwald R, Arca M, and Coleman RA
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- Adult, Aged, Blotting, Western, Cardiomyopathies genetics, Cardiomyopathies metabolism, Chromatography, Thin Layer, Female, HeLa Cells, Humans, Lipase genetics, Male, Microscopy, Fluorescence, Mutagenesis, Site-Directed, Mutation, Missense genetics, Reverse Transcriptase Polymerase Chain Reaction, Fibroblasts metabolism, Lipase metabolism, Lipid Metabolism, Inborn Errors genetics, Lipid Metabolism, Inborn Errors metabolism, Muscular Diseases genetics, Muscular Diseases metabolism, Triglycerides metabolism
- Abstract
The lack of adipose triglyceride lipase (ATGL), a patatin-like phospholipase domain-containing enzyme that hydrolyzes fatty acids from triacylglycerol (TAG) stored in multiple tissues, causes the autosomal recessive disorder neutral lipid storage disease with myopathy (NLSD-M). In two families of Lebanese and Italian origin presenting with NLSD-M, we identified two new missense mutations in highly conserved regions of ATGL (p.Arg221Pro and p.Asn172Lys) and a novel nonsense mutation (p.Trp8X). The Lebanese patients harbor homozygous p.Arg221Pro, whereas the Italian patients are heterozygotes for p.Asn172Lys and the p.Trp8X mutation. The p.Trp8X mutation results in a complete absence of ATGL protein, while the p.Arg221Pro and p.Asn172Lys mutations result in proteins with minimal lipolytic activity. Although these mutations did not affect putative catalytic residues or the lipid droplet (LD)-binding domain of ATGL, cytosolic LDs accumulated in cultured skin fibroblasts from the patients. The missense mutations might destabilize a random coil (p.Asn172Lys) or a helix (p.Arg221Pro) structure within or proximal to the patatin domain of the lipase, thereby interfering with the enzyme activity, while leaving intact the residues required to localize the protein to LDs. Overexpressing wild-type ATGL in one patient's fibroblasts corrected the metabolic defect and effectively reduced the number and area of cellular LDs. Despite the poor lipase activity in vitro, the Lebanese siblings have a mild myopathy and not clinically evident myocardial dysfunction. The patients of Italian origin show a late-onset and slowly progressive skeletal myopathy. These findings suggest that a small amount of correctly localized lipase activity preserves cardiac function in NLSD-M.
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- 2012
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25. Transforming growth factor-beta1 induces microvascular abnormalities through a down-modulation of neural cell adhesion molecule in human hepatocellular carcinoma.
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Balzarini P, Benetti A, Invernici G, Cristini S, Zicari S, Caruso A, Gatta LB, Berenzi A, Imberti L, Zanotti C, Portolani N, Giulini SM, Ferrari M, Ciusani E, Navone SE, Canazza A, Parati EA, and Alessandri G
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- Biomarkers metabolism, Carcinoma, Hepatocellular blood supply, Carcinoma, Hepatocellular pathology, Cell Line, Tumor, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Humans, Immunohistochemistry, Liver Neoplasms blood supply, Liver Neoplasms pathology, Neovascularization, Pathologic, Carcinoma, Hepatocellular metabolism, Down-Regulation, Liver Neoplasms metabolism, Microvessels abnormalities, Neural Cell Adhesion Molecules physiology, Transforming Growth Factor beta1 physiology
- Abstract
Hepatocellular carcinoma (HCC) is a very angiogenic and malignant cancer. Conventional chemotherapy is poorly effective because of the abnormal structural organization of HCC-infiltrating vessels. In previous work, we demonstrated that HCC angiogenesis is driven by transforming growth factor beta-1(TGF-β1)/CD105 axis, stimulating liver-derived microvascular endothelial cells (Ld-MECs) migration. As TGF-β1 also affects mural cells (MCs) recruitment and maturation, we asked whether it may contribute to HCC-induced vascular abnormalities. HCC and adjacent non-neoplastic liver (nNL) biopsies obtained from 12 patients were analyzed by immunohistochemistry for angiogenic markers CD105, TGF-β1, CD44 and vascular endothelial growth factor-a (VEGFa) and for MC markers NG2, α-smooth muscle actin (αSMA) and neural cell adhesion molecule (NCAM). The same markers were also investigated by immunocytochemistry on cultured HCC-derived stromal cells (HCC-StCs) and nNL-derived StCs (nNL-StCs) isolated from the same liver biopsies. Angiogenic factors released by StCs were analyzed by ELISA and the interaction between StCs and Ld-MECs by adhesion assay. Compared with nNL, HCC biopsies showed increased angiogenic markers and αSMA that was localized in vessels. By contrast, NG2 and NCAM were substantially localized in tumor cells but absent in vessels and stroma. Cultured HCC-StCs showed less expression of NG2, αSMA and NCAM. They also demonstrated a lower capacity to release angiogenic factors and adhered on Ld-MECs. HCC-StCs and nNL-StCs treated with TGF-β1 or with of HepG2 (a human hepatoma cell line) derived conditioned medium (CM), down-modulated NCAM expression, whereas anti-NCAM antibodies significantly reduced the adhesion of StCs to Ld-MECs. By further blocking TGF-β1 with anti-TGF-β1 antibodies or with Ly-364947 (a specific inhibitor TGF-β1-receptor) adhesion to Ld-MECs and NCAM expression respectively was partially restored. TGF-β1 contributes to HCC-induced vascular alterations by affecting the interaction between HCC-StCs and Ld-MECs through a down-modulation of NCAM expression.
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- 2012
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26. Three-dimensional self-organizing neural architectures: a neural stem cells reservoir and a system for neurodevelopmental studies.
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Cristini S, Alessandri G, Acerbi F, Ciusani E, Colombo A, Fascio U, Nicosia RF, Invernizzi RW, Gelati M, Parati EA, and Invernici G
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- AC133 Antigen, Antigens, CD metabolism, Axons drug effects, Axons metabolism, Axons ultrastructure, Brain cytology, Brain embryology, CD146 Antigen metabolism, Calcium metabolism, Cell Differentiation drug effects, Cell Proliferation drug effects, Cell Separation, Cells, Cultured, Extracellular Matrix drug effects, Extracellular Matrix metabolism, Fetus cytology, Glutamates pharmacology, Glycoproteins metabolism, Humans, Immunomagnetic Separation, Neural Stem Cells drug effects, Neural Stem Cells metabolism, Neurons drug effects, Neurons ultrastructure, Peptides metabolism, Nervous System cytology, Nervous System growth & development, Neural Stem Cells cytology, Neurons cytology
- Abstract
Complex microenvironmental stimuli influence neural cell properties. To study this, we developed a three-dimensional (3-D) neural culture system, composed of different populations including neurons, astrocytes, and neural stem cells (NSCs). In particular, these last-mentioned cells represent a source potentially exploitable to test drugs, to study neurodevelopment and cell-therapies for neuroregenerations. On seeding on matrigel in a medium supplemented with serum and mitogens, cells obtained from human fetal brain tissue formed 3-D self-organizing neural architectures. Immunocytochemical analysis demonstrated the presence of undifferentiated nestin+ and CD133+ cells, surrounded by β-tub-III+ and GFAP+ cells, suggesting the formation of niches containing potential human NSCs (hNSCs). The presence of hNSCs was confirmed by both neurosphere assay and RT-PCR, and their multipotentiality was demonstrated by both immunofluorescent staining and RT-PCR. Flow cytometry analysis revealed that neurosphere forming cells originating from at least two different subsets expressing, respectively, CD133 and CD146 markers were endowed with different proliferative and differentiation potential. Our data implicate that the complexity of environment within niches and aggregates of heterogeneous neural cell subsets may represent an innovative platform for neurobiological and neurodevelopmental investigations and a reservoir for a rapid expansion of hNSCs.
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- 2011
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27. Dermal fibroblasts display similar phenotypic and differentiation capacity to fat-derived mesenchymal stem cells, but differ in anti-inflammatory and angiogenic potential.
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Blasi A, Martino C, Balducci L, Saldarelli M, Soleti A, Navone SE, Canzi L, Cristini S, Invernici G, Parati EA, and Alessandri G
- Abstract
Background: Mesenchymal stem cells (MSCs) are multipotent stem cells able to differentiate into different cell lineages. However, MSCs represent a subpopulation of a more complex cell composition of stroma cells contained in mesenchymal tissue. Due to a lack of specific markers, it is difficult to distinguish MSCs from other more mature stromal cells such as fibroblasts, which, conversely, are abundant in mesenchymal tissue. In order to find more distinguishing features between MSCs and fibroblasts, we studied the phenotypic and functional features of human adipose-derived MSCs (AD-MSCs) side by side with normal human dermal fibroblasts (HNDFs) in vitro, Methods: AD-MSCs and HNDFs were cultured, expanded and phenotypically characterized by flow cytometry (FC). Immunofluorescence was used to investigate cell differentiation. ELISA assay was used to quantify angiogenic factors and chemokines release. Cultures of endothelial cells (ECs) and a monocyte cell line, U937, were used to test angiogenic and anti-inflammatory properties., Results: Cultured AD-MSCs and HNDFs display similar morphological appearance, growth rate, and phenotypic profile. They both expressed typical mesenchymal markers-CD90, CD29, CD44, CD105 and to a minor extent, the adhesion molecules CD54, CD56, CD106 and CD166. They were negative for the stem cell markers CD34, CD146, CD133, CD117. Only aldehyde dehydrogenase (ALDH) was expressed. Neither AD-MSCs nor HNDFs differed in their multi-lineage differentiation capacity; they both differentiated into osteoblast, adipocyte, and also into cardiomyocyte-like cells. In contrast, AD-MSCs, but not HNDFs, displayed strong angiogenic and anti-inflammatory activity. AD-MSCs released significant amounts of VEGF, HGF and Angiopoietins and their conditioned medium (CM) stimulated ECs proliferation and tube formations. In addition, CM-derived AD-MSCs (AD-MSCs-CM) inhibited adhesion molecules expression on U937 and release of RANTES and MCP-1. Finally, after priming with TNFα, AD-MSCs enhanced their anti-inflammatory potential; while HNDFs acquired pro-inflammatory activity., Conclusions: AD-MSCs cannot be distinguished from HNDFs in vitro by evaluating their phenotypic profile or differentiation potential, but only through the analysis of their anti-inflammatory and angiogenic properties. These results underline the importance of evaluating the angiogenic and anti-inflammatory features of MSCs preparation. Their priming with inflammatory cytokines prior to transplantation may improve their efficacy in cell-based therapies for tissue regeneration.
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- 2011
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28. Nanotechnology advances in brain tumors: the state of the art.
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Invernici G, Cristini S, Alessandri G, Navone SE, Canzi L, Tavian D, Redaelli C, Acerbi F, and Parati EA
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- Animals, Brain Neoplasms diagnosis, Carcinoma diagnosis, Diagnostic Imaging methods, Diagnostic Imaging trends, Drug Carriers chemical synthesis, Drug Carriers chemistry, Drug Delivery Systems methods, Drug Delivery Systems trends, Genetic Therapy methods, Humans, Molecular Targeted Therapy methods, Molecular Targeted Therapy trends, Nanoparticles therapeutic use, Nanotechnology methods, Brain Neoplasms therapy, Carcinoma therapy, Nanotechnology trends
- Abstract
Primary malignant central nervous system (CNS) tumors only represent about 2% of all cancers. However, they are very often associated with high morbidity and mortality. Despite current standard-of-care therapy, such as surgery, irradiation, and chemotherapy, neither cure nor any toxic therapy against malignant CNS tumors has been developed so far. Nanotechnology may alter this situation. It offers a new promise for cancer diagnosis and treatment. This emerging technology, by developing and manufacturing materials using atomic and molecular elements, can provide a platform for the combination of diagnostics, therapeutics and delivery to the tumor, with subsequent monitoring of the response. This review focuses on recent developments in cancer nanotechnology with particular attention to nanoparticle systems, important tools for the improvement of drug delivery in brain tumor. The latest advances in both the research sector and in recent patents for cancer imaging and therapy are discussed.
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- 2011
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29. Mesenchymal stromal cells primed with paclitaxel provide a new approach for cancer therapy.
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Pessina A, Bonomi A, Coccè V, Invernici G, Navone S, Cavicchini L, Sisto F, Ferrari M, Viganò L, Locatelli A, Ciusani E, Cappelletti G, Cartelli D, Arnaldo C, Parati E, Marfia G, Pallini R, Falchetti ML, and Alessandri G
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- ATP Binding Cassette Transporter, Subfamily B, Member 1 metabolism, Animals, Antineoplastic Agents metabolism, Antineoplastic Agents therapeutic use, Biological Transport, Cell Line, Tumor, Cell Proliferation drug effects, Endothelial Cells drug effects, Endothelial Cells pathology, Gene Expression Regulation, Neoplastic drug effects, Humans, Kinetics, Mesenchymal Stem Cells metabolism, Mesenchymal Stem Cells pathology, Mice, Neoplasms blood supply, Neoplasms metabolism, Neovascularization, Pathologic drug therapy, Paclitaxel metabolism, Paclitaxel therapeutic use, Tumor Microenvironment drug effects, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Mesenchymal Stem Cells drug effects, Neoplasms drug therapy, Neoplasms pathology, Paclitaxel pharmacology
- Abstract
Background: Mesenchymal stromal cells may represent an ideal candidate to deliver anti-cancer drugs. In a previous study, we demonstrated that exposure of mouse bone marrow derived stromal cells to Doxorubicin led them to acquire anti-proliferative potential towards co-cultured haematopoietic stem cells (HSCs). We thus hypothesized whether freshly isolated human bone marrow Mesenchymal stem cells (hMSCs) and mature murine stromal cells (SR4987 line) primed in vitro with anti-cancer drugs and then localized near cancer cells, could inhibit proliferation., Methods and Principal Findings: Paclitaxel (PTX) was used to prime culture of hMSCs and SR4987. Incorporation of PTX into hMSCs was studied by using FICT-labelled-PTX and analyzed by FACS and confocal microscopy. Release of PTX in culture medium by PTX primed hMSCs (hMSCsPTX) was investigated by HPLC. Culture of Endothelial cells (ECs) and aorta ring assay were used to test the anti-angiogenic activity of hMSCsPTX and PTX primed SR4987(SR4987PTX), while anti-tumor activity was tested in vitro on the proliferation of different tumor cell lines and in vivo by co-transplanting hMSCsPTX and SR4987PTX with cancer cells in mice. Nevertheless, despite a loss of cells due to chemo-induced apoptosis, both hMSCs and SR4987 were able to rapidly incorporate PTX and could slowly release PTX in the culture medium in a time dependent manner. PTX primed cells acquired a potent anti-tumor and anti-angiogenic activity in vitro that was dose dependent, and demonstrable by using their conditioned medium or by co-culture assay. Finally, hMSCsPTX and SR4987PTX co-injected with human cancer cells (DU145 and U87MG) and mouse melanoma cells (B16) in immunodeficient and in syngenic mice significantly delayed tumor takes and reduced tumor growth., Conclusions: These data demonstrate, for the first time, that without any genetic manipulation, mesenchymal stromal cells can uptake and subsequently slowly release PTX. This may lead to potential new tools to increase efficacy of cancer therapy.
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- 2011
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30. Tumour vascularization via endothelial differentiation of glioblastoma stem-like cells.
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Ricci-Vitiani L, Pallini R, Biffoni M, Todaro M, Invernici G, Cenci T, Maira G, Parati EA, Stassi G, Larocca LM, and De Maria R
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- Animals, Biomarkers, Tumor metabolism, Cell Line, Tumor, Cell Lineage, Chromosome Aberrations, Endothelial Cells metabolism, Glioblastoma genetics, Humans, In Situ Hybridization, Fluorescence, Mice, Mice, Inbred NOD, Mice, SCID, Mice, Transgenic, Models, Biological, Neoplasm Transplantation pathology, Neovascularization, Pathologic genetics, Neural Stem Cells metabolism, Transplantation, Heterologous pathology, Cell Differentiation, Endothelial Cells pathology, Endothelium, Vascular pathology, Glioblastoma blood supply, Glioblastoma pathology, Neovascularization, Pathologic pathology, Neural Stem Cells pathology
- Abstract
Glioblastoma is a highly angiogenetic malignancy, the neoformed vessels of which are thought to arise by sprouting of pre-existing brain capillaries. The recent demonstration that a population of glioblastoma stem-like cells (GSCs) maintains glioblastomas indicates that the progeny of these cells may not be confined to the neural lineage. Normal neural stem cells are able to differentiate into functional endothelial cells. The connection between neural stem cells and the endothelial compartment seems to be critical in glioblastoma, where cancer stem cells closely interact with the vascular niche and promote angiogenesis through the release of vascular endothelial growth factor (VEGF) and stromal-derived factor 1 (refs 5-9). Here we show that a variable number (range 20-90%, mean 60.7%) of endothelial cells in glioblastoma carry the same genomic alteration as tumour cells, indicating that a significant portion of the vascular endothelium has a neoplastic origin. The vascular endothelium contained a subset of tumorigenic cells that produced highly vascularized anaplastic tumours with areas of vasculogenic mimicry in immunocompromised mice. In vitro culture of GSCs in endothelial conditions generated progeny with phenotypic and functional features of endothelial cells. Likewise, orthotopic or subcutaneous injection of GSCs in immunocompromised mice produced tumour xenografts, the vessels of which were primarily composed of human endothelial cells. Selective targeting of endothelial cells generated by GSCs in mouse xenografts resulted in tumour reduction and degeneration, indicating the functional relevance of the GSC-derived endothelial vessels. These findings describe a new mechanism for tumour vasculogenesis and may explain the presence of cancer-derived endothelial-like cells in several malignancies.
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- 2010
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31. Neural precursor-derived astrocytes of wobbler mice induce apoptotic death of motor neurons through reduced glutamate uptake.
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Diana V, Ottolina A, Botti F, Fumagalli E, Calcagno E, De Paola M, Cagnotto A, Invernici G, Parati E, Curti D, and Mennini T
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- Animals, Cell Communication physiology, Cell Death physiology, Cells, Cultured, Coculture Techniques, Mice, Mice, Neurologic Mutants, Motor Neurons metabolism, Nerve Degeneration metabolism, Nerve Degeneration pathology, Apoptosis physiology, Astrocytes metabolism, Astrocytes pathology, Glutamic Acid metabolism, Motor Neurons pathology, Stem Cells metabolism, Stem Cells pathology
- Abstract
In the present study, we investigated whether cultured astrocytes derived from adult neural precursor cells (NPCs) obtained from the subventricular zone (SVZ) of wobbler mice display metabolic traits of the wobbler astrocytes in situ and in primary culture. We also utilized NPC-derived astrocytes as a tool to investigate the involvement of astrocytes in the molecular mechanism of MND focusing on the possible alteration of glutamate reuptake since excitotoxicity glutamate-mediated may be a contributory pathway. NPC-derived wobbler astrocytes are characterized by high immunoreactivity for GFAP, significant decrease of glutamate uptake and reduced immunoreactivity for glutamate transporters GLT1 and GLAST. Spinal cord motor neurons obtained from healthy mouse embryos, when co-cultured with wobbler NPC-derived astrocytes, show reduced viability and morphologic alterations. These suffering motor neurons are caspase-7 positive, and treatment with anti-apoptotic drug V5 increases cell survival. Physical contact with wobbler astrocytes is not essential because purified motor neurons display reduced survival also when treated with the medium conditioned by wobbler NPC-derived astrocytes. Toxic levels of glutamate were revealed by HPLC assay in the extracellular medium of wobbler NPC-derived astrocytes, whereas the level of intracellular glutamate is reduced if compared with controls. Moreover, glutamate receptor antagonists are able to enhance motor neuron survival. Therefore, our results demonstrate that astrocytes derived from wobbler neural precursor cells display impaired glutamate homeostasis that may play a crucial role in motor neuron degeneration. Finally, the cultured astrocytes derived from NPCs of adult mice may offer a useful alternative in vitro model to study the molecular mechanisms involved in neurodegeneration., (Copyright 2010 Elsevier Inc. All rights reserved.)
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- 2010
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32. Omentum-derived stromal cells improve myocardial regeneration in pig post-infarcted heart through a potent paracrine mechanism.
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De Siena R, Balducci L, Blasi A, Montanaro MG, Saldarelli M, Saponaro V, Martino C, Logrieco G, Soleti A, Fiobellot S, Madeddu P, Rossi G, Ribatti D, Crovace A, Cristini S, Invernici G, Parati EA, and Alessandri G
- Subjects
- Animals, Apoptosis, Cell Differentiation, Cell Proliferation, Cells, Cultured, Endothelial Cells pathology, Endothelial Cells physiology, Female, Heart physiology, Humans, In Vitro Techniques, Mice, Myocardial Infarction pathology, Myocardial Infarction physiopathology, Myocytes, Cardiac pathology, Myocytes, Cardiac physiology, Neovascularization, Physiologic, Paracrine Communication, Stromal Cells cytology, Swine, Myocardial Infarction therapy, Omentum cytology, Regeneration physiology, Stromal Cells physiology, Stromal Cells transplantation
- Abstract
Cell-based therapy could be a valid option to treat myocardial infarct (MI). Adipose-derived stromal cells (ADStCs) have demonstrated tissue regenerative potential including cardiomyogenesis. Omentum is an extremely rich source of visceral fat and its accumulation seems to correlate with cardiovascular diseases. We investigated the capacity of human fat Omentum-derived StCs (FOStCs) to affect heart function upon acute infarct in pigs induced by permanent ligation of the anterior interventricular artery (IVA). We demonstrated for the first time that the local injection of 50x10(6) of FOStCs ameliorates the functional parameters of post-infarct heart. Most importantly, histology of FOStCs treated hearts demonstrated a substantial improvement of cardiomyogenesis. In culture, FOStCs produced an impressive number and amount of angiogenic factors and cytokines. Moreover, the conditioned medium of FOStCs (FOStCs-CM) stimulates in vitro cardiac endothelial cells (ECs) proliferation and vascular morphogenesis and inhibits monocytes, EC activation and cardiomyocyte apoptosis. Since FOStCs in vivo did not trans-differentiate into cardiomyocyte-like cells, we conclude that FOStCs efficacy was presumably mediated by a potent paracrine mechanism involving molecules that concomitantly improved angiogenesis, reduced inflammation and prevented cardiomyocytes death. Our results highlight for the first time the important role that human FOStCs may have in cardiac regeneration.
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- 2010
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33. Human neural stem cells: a model system for the study of Lesch-Nyhan disease neurological aspects.
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Cristini S, Navone S, Canzi L, Acerbi F, Ciusani E, Hladnik U, de Gemmis P, Alessandri G, Colombo A, Parati E, and Invernici G
- Subjects
- Biomarkers metabolism, Cell Differentiation genetics, Dopamine metabolism, Fluorescent Antibody Technique, Gene Expression Profiling, Gene Expression Regulation, Humans, Lesch-Nyhan Syndrome genetics, Reverse Transcriptase Polymerase Chain Reaction, Lesch-Nyhan Syndrome pathology, Models, Biological, Neurons metabolism, Stem Cells metabolism
- Abstract
The study of Lesch-Nyhan-diseased (LND) human brain is crucial for understanding how mutant hypoxanthine-phosphoribosyltransferase (HPRT) might lead to neuronal dysfunction. Since LND is a rare, inherited disorder caused by a deficiency of the enzyme HPRT, human neural stem cells (hNSCs) that carry this mutation are a precious source for delineating the consequences of HPRT deficiency and for developing new treatments. In our study we have examined the effect of HPRT deficiency on the differentiation of neurons in hNSCs isolated from human LND fetal brain. We have examined the expression of a number of transcription factors essential for neuronal differentiation and marker genes involved in dopamine (DA) biosynthetic pathway. LND hNSCs demonstrate aberrant expression of several transcription factors and DA markers. HPRT-deficient dopaminergic neurons also demonstrate a striking deficit in neurite outgrowth. These results represent direct experimental evidence for aberrant neurogenesis in LND hNSCs and suggest developmental roles for other housekeeping genes in neurodevelopmental disease. Moreover, exposure of the LND hNSCs to retinoic acid medium elicited the generation of dopaminergic neurons. The lack of precise understanding of the neurological dysfunction in LND has precluded development of useful therapies. These results evidence aberrant neurogenesis in LND hNSCs and suggest a role for HPRT gene in neurodevelopment. These cells combine the peculiarity of a neurodevelopmental model and a human, neural origin to provide an important tool to investigate the pathophysiology of HPRT deficiency and more broadly demonstrate the utility of human neural stem cells for studying the disease and identifying potential therapeutics.
- Published
- 2010
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34. Stem cell patents: an innovative approach to anti-cancer drug discovery.
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Navone S, Cristini S, Canzi L, Parati EA, and Invernici G
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- Animals, Humans, Neoplasms drug therapy, Neoplastic Stem Cells metabolism, Stem Cell Transplantation methods, Antineoplastic Agents therapeutic use, Drug Discovery ethics, Drug Discovery legislation & jurisprudence, Neoplasms therapy, Patents as Topic, Stem Cell Transplantation legislation & jurisprudence, Stem Cells physiology
- Abstract
Over the last decade, improvements in cancer therapies have prolonged the lives of cancer patients. Despite dramatic advances in imaging technology, surgical techniques, and adjuvant radio- and chemotherapy, the overall prognosis of this disease remains dismal. In light of this, there is an urgent need for the development of more effective therapies that can target residual disseminated tumor burden. Given the heterogeneity of tumors in general, no one strategy is likely to provide a satisfactory treatment regimen. Until the middle of the 20th century, medical treatments were limited to options like drugs, surgery, antibiotics, and radiation, but in the last years stem cells, due to their pathotropism, have become particularly attractive candidates not only to replace damaged tissue in degenerative pathologies, but also to deliver therapeutic molecules in patients with disseminated metastatic cancer. Worldwide there have been over 2000 patent applications involving human and non-human stem cells, of which one quarter refer to embryonic stem cells. Over one third of all stem cell applications and one quarter of all embryonic stem cell applications have been granted. The aim of this review is primarily to focus on the recent development of stem cell patents in cancer treatments.
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- 2010
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35. Human CD133+ progenitor cells promote the healing of diabetic ischemic ulcers by paracrine stimulation of angiogenesis and activation of Wnt signaling.
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Barcelos LS, Duplaa C, Kränkel N, Graiani G, Invernici G, Katare R, Siragusa M, Meloni M, Campesi I, Monica M, Simm A, Campagnolo P, Mangialardi G, Stevanato L, Alessandri G, Emanueli C, and Madeddu P
- Subjects
- AC133 Antigen, Animals, Antigens, CD analysis, Aorta embryology, Cell Differentiation, Cell Movement, Cell Proliferation, Cell Survival, Cells, Cultured, Culture Media, Conditioned metabolism, Diabetes Mellitus, Experimental metabolism, Diabetes Mellitus, Experimental physiopathology, Diabetes Mellitus, Experimental surgery, Diabetic Foot etiology, Diabetic Foot metabolism, Diabetic Foot physiopathology, Fetal Stem Cells immunology, Fetal Stem Cells metabolism, Glycoproteins analysis, Humans, Intercellular Signaling Peptides and Proteins metabolism, Interleukin-8 metabolism, Ischemia metabolism, Ischemia physiopathology, Ischemia surgery, Male, Membrane Proteins metabolism, Mice, Paracrine Communication, Peptides analysis, Signal Transduction, Time Factors, Vascular Endothelial Growth Factor A metabolism, Diabetes Mellitus, Experimental complications, Diabetic Foot surgery, Fetal Stem Cells transplantation, Ischemia complications, Lower Extremity blood supply, Neovascularization, Physiologic, Stem Cell Transplantation, Wnt Proteins metabolism, Wound Healing
- Abstract
We evaluated the healing potential of human fetal aorta-derived CD133(+) progenitor cells and their conditioned medium (CD133(+) CCM) in a new model of ischemic diabetic ulcer. Streptozotocin-induced diabetic mice underwent bilateral limb ischemia and wounding. One wound was covered with collagen containing 2x10(4) CD133(+) or CD133(-) cells or vehicle. The contralateral wound, covered with only collagen, served as control. Fetal CD133(+) cells expressed high levels of wingless (Wnt) genes, which were downregulated following differentiation into CD133(-) cells along with upregulation of Wnt antagonists secreted frizzled-related protein (sFRP)-1, -3, and -4. CD133(+) cells accelerated wound closure as compared with CD133(-) or vehicle and promoted angiogenesis through stimulation of endothelial cell proliferation, migration, and survival by paracrine effects. CD133(+) cells secreted high levels of vascular endothelial growth factor (VEGF)-A and interleukin (IL)-8. Consistently, CD133(+) CCM accelerated wound closure and reparative angiogenesis, with this action abrogated by co-administering the Wnt antagonist sFRP-1 or neutralizing antibodies against VEGF-A or IL-8. In vitro, these effects were recapitulated following exposure of high-glucose-primed human umbilical vein endothelial cells to CD133(+) CCM, resulting in stimulation of migration, angiogenesis-like network formation and induction of Wnt expression. The promigratory and proangiogenic effect of CD133(+) CCM was blunted by sFRP-1, as well as antibodies against VEGF-A or IL-8. CD133(+) cells stimulate wound healing by paracrine mechanisms that activate Wnt signaling pathway in recipients. These preclinical findings open new perspectives for the cure of diabetic ulcers.
- Published
- 2009
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36. Transforming growth factor-beta1 and CD105 promote the migration of hepatocellular carcinoma-derived endothelium.
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Benetti A, Berenzi A, Gambarotti M, Garrafa E, Gelati M, Dessy E, Portolani N, Piardi T, Giulini SM, Caruso A, Invernici G, Parati EA, Nicosia R, and Alessandri G
- Subjects
- Antigens, CD analysis, Cadherins analysis, Carcinoma, Hepatocellular pathology, Cell Line, Tumor, Cell Movement, Endoglin, Humans, Hyaluronan Receptors analysis, Immunohistochemistry, Liver Neoplasms pathology, Receptors, Cell Surface analysis, Transforming Growth Factor beta1 analysis, Antigens, CD physiology, Carcinoma, Hepatocellular blood supply, Endothelial Cells physiology, Liver Neoplasms blood supply, Neovascularization, Pathologic etiology, Receptors, Cell Surface physiology, Transforming Growth Factor beta1 physiology
- Abstract
Hepatocellular carcinoma (HCC) is one of most malignant and aggressive human tumors. Transforming growth factor-beta1 (TGF-beta1) and its coreceptor CD105 have been shown to contribute to HCC malignant progression. TGF-beta1 and CD105 have also been implicated in angiogenesis, but their role in the vascularization of HCC has not been investigated. To fill this gap, we studied the effect of TGF-beta1 and CD105 on HCC-derived endothelium. By using immunomagnetic beads, we isolated and cultured endothelial cells (ECs) from HCC (HCC-EC) and adjacent nonneoplastic tissue (nNL-ECs) obtained from 24 liver biopsies. HCC and nNL biopsies were also analyzed by immunohistochemistry for the expression of CD105, TGF-beta1, Ve-cadherin (Ve-cad), CD44, beta-catenin, and E-cadherin. Compared with nNL-ECs, HCC-ECs had higher expression of CD105, enhanced spontaneous motility, and greater capacity to migrate in response to TGF-beta1 (5 ng/mL), particularly in the presence of a fibronectin matrix. The chemotactic effect of TGF-beta1 was blocked by anti-CD105 antibodies and correlated with the grade of HCC malignancy. Histologic examination of HCC biopsies showed that HCCs with the worse malignant features had the highest expression of TGF-beta1, CD105, and angiogenic markers (Ve-cad and CD44). Because CD105 was highly expressed in microvessels at the tumor periphery and TGF-beta1 staining was only found in neoplastic hepatocytes, we conclude that HCC-derived TGF-beta1 may act as a chemoattractant for CD105-expressing ECs and as a promoter of tumor angiogenesis. Thus, drugs that selectively target the TGF-beta1/CD105 axis may interfere with HCC-related angiogenesis and HCC progression.
- Published
- 2008
- Full Text
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37. Human fetal aorta-derived vascular progenitor cells: identification and potential application in ischemic diseases.
- Author
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Invernici G, Madeddu P, Emanueli C, Parati EA, and Alessandri G
- Abstract
Vasculogenesis, the formation of blood vessels in embryonic or fetal tissue mediated by immature vascular cells (i.e., angioblasts), is poorly understood. Here we report a summary of our recent studies on the identification of a population of vascular progenitor cells (VPCs) in human fetal aorta. These undifferentiated mesenchymal cells co-express endothelial and myogenic markers (CD133+, CD34+, KDR+, desmin+) and are localized in outer layer of the aortic stroma of 11-12 weeks old human fetuses. Under stimulation with VEGF-A or PDGF-BB, VPCs give origin to a mixed population of mature endothelial and mural cells, respectively. When embedded in a three-dimensional collagen gel, VPCs organize into cohesive cellular cords that resembled mature vascular structures. The therapeutic efficacy of a small number of VPCs transplanted into ischemic limb muscle was demonstrated in immunodeficient mice. Investigation of the effect of VPCs on experimental heart ischemia and on diabetic ischemic ulcers in mice is in progress and seems to confirm their efficacy. On the whole, fetal aorta represents an important source for the investigation of phenotypic and functional features of human vascular progenitor cells.
- Published
- 2008
- Full Text
- View/download PDF
38. Human adult skeletal muscle stem cells differentiate into cardiomyocyte phenotype in vitro.
- Author
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Invernici G, Cristini S, Madeddu P, Brock S, Spillmann F, Bernasconi P, Cappelletti C, Calatozzolo C, Fascio U, Bisleri G, Muneretto C, Alessandri G, and Parati EA
- Subjects
- Adult, Aged, Animals, Becaplermin, Humans, Mice, Mice, Inbred Strains, Middle Aged, Muscle, Skeletal metabolism, Myocardial Ischemia metabolism, Myocytes, Cardiac metabolism, Phenotype, Platelet-Derived Growth Factor pharmacology, Proto-Oncogene Proteins c-sis, Stem Cells metabolism, Tretinoin pharmacology, Cell Differentiation, Muscle, Skeletal cytology, Myocytes, Cardiac cytology, Stem Cells cytology
- Abstract
Cell transplantation to repair or regenerate injured myocardium is a new frontier in the treatment of cardiovascular disease. Most studies on stem cell transplantation therapy in both experimental heart infarct and in phase-I human clinical trials have focused on the use of undifferentiated stem cells. Based on our previous observations demonstrating the presence of multipotent progenitor cells in human adult skeletal muscle, in this study we investigated the capacity of these progenitors to differentiate into cardiomyocytes. Here we show an efficient protocol for the cardiomyogenic differentiation of human adult skeletal muscle stem cells in vitro. We found that treatment with Retinoic Acid directed cardiomyogenic differentiation of skeletal muscle stem cells in vitro. After Retinoic Acid treatment, cells expressed cardiomyocyte markers and acquired spontaneous contraction. Functional assays exhibited cardiac-like response to increased extracellular calcium. When cocultured with mouse cardiomyocytes, Retinoic Acid-treated skeletal muscle stem cells expressed connexin43 and when transplanted into ischemic heart were detectable even 5 weeks after injection. Based on these results, we can conclude that human adult skeletal muscle stem cells, if opportunely treated, can transdifferentiate into cells of cardiac lineage and once injected into infarcted heart can integrate, survive in cardiac tissue and improve the cardiac function.
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- 2008
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39. Human fetal aorta contains vascular progenitor cells capable of inducing vasculogenesis, angiogenesis, and myogenesis in vitro and in a murine model of peripheral ischemia.
- Author
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Invernici G, Emanueli C, Madeddu P, Cristini S, Gadau S, Benetti A, Ciusani E, Stassi G, Siragusa M, Nicosia R, Peschle C, Fascio U, Colombo A, Rizzuti T, Parati E, and Alessandri G
- Subjects
- AC133 Antigen, Angiopoietin-2 genetics, Angiopoietin-2 metabolism, Animals, Antigens, CD metabolism, Antigens, CD34 metabolism, Aorta metabolism, Becaplermin, Biomarkers metabolism, Blood Vessels cytology, Blood Vessels embryology, Cell Lineage, Cells, Cultured, Glycoproteins metabolism, Humans, Mice, Peptides metabolism, Platelet-Derived Growth Factor metabolism, Proto-Oncogene Proteins c-sis, Vascular Endothelial Growth Factor A genetics, Vascular Endothelial Growth Factor A metabolism, Vascular Endothelial Growth Factor Receptor-2 metabolism, Aorta cytology, Aorta embryology, Fetus anatomy & histology, Ischemia, Muscle Development physiology, Neovascularization, Physiologic, Stem Cells physiology
- Abstract
Vasculogenesis, the formation of blood vessels in embryonic or fetal tissue mediated by immature vascular cells (ie, angioblasts), is poorly understood. We report the identification of a population of vascular progenitor cells (hVPCs) in the human fetal aorta composed of undifferentiated mesenchymal cells that coexpress endothelial and myogenic markers. Under culture conditions that promoted cell differentiation, hVPCs gave rise to a mixed population of mature endothelial and mural cells when progenitor cells were stimulated with vascular endothelial growth factor-A or platelet-derived growth factor-betabeta. hVPCs grew as nonadherent cells and, when embedded in a three-dimensional collagen gel, reorganized into cohesive cellular cords that resembled mature vascular structures. hVPC-conditioned medium contained angiogenic substances (vascular endothelial growth factor-A and angiopoietin-2) and strongly stimulated the proliferation of endothelial cells. We also demonstrate the therapeutic efficacy of a small number of hVPCs transplanted into ischemic limb muscle of immunodeficient mice. hVPCs markedly improved neovascularization and inhibited the loss of endogenous endothelial cells and myocytes, thus ameliorating the clinical outcome from ischemia. We conclude that fetal aorta represents an important source for the investigation of the phenotypic and functional features of human vascular progenitor cells.
- Published
- 2007
- Full Text
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40. Melanoma contains CD133 and ABCG2 positive cells with enhanced tumourigenic potential.
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Monzani E, Facchetti F, Galmozzi E, Corsini E, Benetti A, Cavazzin C, Gritti A, Piccinini A, Porro D, Santinami M, Invernici G, Parati E, Alessandri G, and La Porta CA
- Subjects
- AC133 Antigen, ATP Binding Cassette Transporter, Subfamily G, Member 2, Animals, Biomarkers metabolism, Blotting, Western, Immunohistochemistry methods, Mice, Mice, Inbred NOD, Mice, SCID, Neovascularization, Pathologic, Reverse Transcriptase Polymerase Chain Reaction methods, Transplantation, Heterologous, Tumor Cells, Cultured, ATP-Binding Cassette Transporters metabolism, Antigens, CD metabolism, Glycoproteins metabolism, Melanoma metabolism, Neoplasm Proteins metabolism, Peptides metabolism, Skin Neoplasms metabolism
- Abstract
The failure to eradicate most cancers and in particular melanoma may be as fundamental as a misidentification of the target. The identification of cancer stem/initiating cells within the tumour population with a crucial role for tumour formation may open new pharmacological perspectives. Our data show three main novelties for human melanoma: firstly, melanoma biopsy contains a subset of cells expressing CD133 (CD133+) and the latter is able to develop a Mart-1 positive tumour in NOD-SCID mice. Secondly, the WM115, a human melanoma cell line, has been found to express both CD133 and ABCG2 markers. This cell line grows as floating spheroids, expresses typical progenitors and mature neuronal/oligodendrocyte markers and is able to transdifferentiate into astrocytes or mesenchymal lineages under specific growth conditions. As in xenografts generated with CD133+ biopsy melanoma cells, those produced by the cell line displayed lower levels of CD133 and ABCG2. Thirdly, the WM115 cells express the most important angiogenic and lymphoangiogenic factors such as notch 4, prox1 and podoplanin which can cooperate in the development of the tumourigenic capability of melanoma in vivo. Therefore, in this study, we demonstrate the presence of stem/initiating subsets in melanoma both in biopsy and in an established melanoma cell line grown in vitro and in xenografts. Interestingly, considering that melanoma gives metastasis primarily through lymphatic vessels, herein, we demonstrated that a melanoma cell line expresses typical lymphoangiogenic factors.
- Published
- 2007
- Full Text
- View/download PDF
41. Isolation and culture of human muscle-derived stem cells able to differentiate into myogenic and neurogenic cell lineages.
- Author
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Alessandri G, Pagano S, Bez A, Benetti A, Pozzi S, Iannolo G, Baronio M, Invernici G, Caruso A, Muneretto C, Bisleri G, and Parati E
- Subjects
- Actins analysis, Aged, Animals, Cell Culture Techniques methods, Cell Division drug effects, Cell Lineage, Cell Separation methods, Clone Cells, Desmin analysis, Female, Growth Substances pharmacology, Humans, Immunohistochemistry, Male, Middle Aged, Muscle Proteins analysis, Muscle, Skeletal chemistry, Nerve Tissue Proteins analysis, Pluripotent Stem Cells chemistry, Rats, Rats, Sprague-Dawley, Spinal Cord, Stem Cell Transplantation, Vimentin analysis, Cell Differentiation, Muscle, Skeletal cytology, Neurons cytology, Pluripotent Stem Cells cytology
- Abstract
Background: Skeletal-muscle-derived stem cells seem to be a distinct population of immature progenitors of satellite cells, but their functional properties remain unclear, especially in human adult tissue. We investigated their differentiation in samples of skeletal muscle obtained from adults undergoing cardiovascular surgery., Methods: Samples were obtained from the brachioradialis muscle of 12 patients in whom the radial artery was the conduit for myocardial revascularisation. The stem cells were isolated by a procedure similar to that used for rat gastrocnemius and cultured in medium optimised for growth of neural stem cells. Cytometry was used for phenotypic characterisation and immunocytochemistry and RT-PCR to assess differentiation. Immunohistochemistry was used to examine engraftment of skeletal-muscle-derived stem cells into injured rat spinal cord., Findings: The skeletal-muscle stem cells consisted of two distinct types: one with the typical spindle morphology of satellite cells, the other of rounded cells. Some cultures could be maintained for longer than 6 months. The cells were mainly positive for desmin and to a lesser extent CD105, vimentin, and AC133/CD133, but negative for FLK-1/KDR, CD34, CD31, CD45, von Willebrand factor, Ve-cadherins, and BCL2. After in-vitro differentiation, the cells were able to organise skeletal-muscle fibres and stained positively for striated-muscle actin, smooth-muscle actin, and desmin. Moreover, they differentiated into astrocytes and neurons, as confirmed by positive staining for characteristic proteins., Interpretation: Adult human skeletal muscle includes a population of progenitor stem cells that can generate cells of the same lineage and cells with neurogenic properties. Muscle may therefore be a tissue source for the isolation of pluripotent stem cells for development of cell-based therapies for human myogenic and neurogenic diseases.
- Published
- 2004
- Full Text
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