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149 results on '"Martens, A.C.M."'

2. Mesenchymal stem cell therapy in proteoglycan induced arthritis

Author

Swart, J. F., de Roock, S., Hofhuis, F. M., Rozemuller, H., van den Broek, T., Moerer, P., Broere, F., van Wijk, F., Kuis, W., Prakken, B. J., Martens, a.c.m, Wulffraat, N. M., Infection & Immunity, Risk Assessment, dI&I RA-I&I I&I, and LS Immunologie

Subjects
Rheumatology, Biochemistry, Genetics and Molecular Biology(all), Immunology, Immunology and Allergy
Abstract

Objectives: To explore the immunosuppressive effect and mechanism of action of intraperitoneal (ip) and intra-articular (ia) mesenchymal stem cell (MSC) injection in proteoglycan induced arthritis (PGIA). Methods: MSC were administered ip or ia after establishment of arthritis. We used serial bioluminescence imaging (BLI) to trace luciferase-transfected MSC. Mice were sacrificed at different time points to examine immunomodulatory changes in blood and secondary lymphoid organs. Results: Both ip and local ia MSC injection resulted in a beneficial clinical and histological effect on established PGIA. BLI showed that MSC ip and ia in arthritic mice are largely retained for several weeks in the peritoneal cavity or injected joint respectively, without signs of migration. Following MSC treatment pathogenic PG-specific IgG2a antibodies in serum decreased. The Th2 cytokine IL-4 was only upregulated in PG-stimulated lymphocytes from spleens in ip treated mice and in lymphocytes from draining lymph nodes in ia treated mice. An increase in production of IL-10 was seen with equal distribution. Although IFN-? was also elevated, the IFN-?/IL-4 ratio in MSC treated mice was opposite to the ratio in (untreated) active PGIA. Conclusions: MSC treatment, both ip and ia, suppresses PGIA, a non-collagen induced arthritis model. MSC are largely retained for weeks in the injection region. MSC treatment induced at the region of injection a deviation of PG-specific immune responses, suggesting a more regulatory phenotype with production of IL-4 and IL-10, but also of IFN-γ, and a systemic decrease of pathogenic PG-specific IgG2a antibodies. These findings underpin the potential of MSC treatment in resistant arthritis.

Published
2015

3. Non-canonical PRC1.1 Targets Active Genes Independent of H3K27me3 and Is Essential for Leukemogenesis

Author

van den Boom, V., Maat, H., Geugien, M., Lopez, A.R., Sotoca, A.M., Jaques, J., Brouwers-Vos, A.Z., Fusetti, F., Groen, R.W.J., Yuan, H.P., Martens, A.C.M., Stunnenberg, H.G., Vellenga, E., Martens, J.H.A., Schuringa, J.J., van den Boom, V., Maat, H., Geugien, M., Lopez, A.R., Sotoca, A.M., Jaques, J., Brouwers-Vos, A.Z., Fusetti, F., Groen, R.W.J., Yuan, H.P., Martens, A.C.M., Stunnenberg, H.G., Vellenga, E., Martens, J.H.A., and Schuringa, J.J.

Abstract

Contains fulltext : 156956.pdf (publisher's version ) (Open Access)

Published
2016

5. In vitro induction of alkaline phosphatase levels predicts in vivo bone forming capacity of human bone marrow stromal cells

Author

Prins, H.J., Braat, A.K., Gawlitta, D., Dhert, W.J.A., Egan, D.A., Tijssen-Slump, E., Yuan, H., Coffer, P.J, Rozemuller, H., Martens, A.C.M., Dep Gezondheidszorg Paard, Geneeskunde van gezelschapsdieren, CSCA TR1, Tissue Repair, Faculty of Science and Technology, Dep Gezondheidszorg Paard, Geneeskunde van gezelschapsdieren, CSCA TR1, Tissue Repair, and CCA - Innovative therapy

Subjects
Stromal cell, Population, Gene Expression, Bone Marrow Cells, Biology, Mice, In vivo, Osteogenesis, medicine, Animals, Humans, education, Child, lcsh:QH301-705.5, Cells, Cultured, Medicine(all), education.field_of_study, Tissue Engineering, IR-88599, Cell Differentiation, Mesenchymal Stem Cells, General Medicine, Cell Biology, Chondrogenesis, Alkaline Phosphatase, In vitro, Cell biology, METIS-300672, medicine.anatomical_structure, lcsh:Biology (General), Adipogenesis, Enzyme Induction, Immunology, Alkaline phosphatase, Bone marrow, Developmental Biology
Abstract

One of the applications of bone marrow stromal cells (BMSCs) that are produced by ex vivo expansion is for use in in vivo bone tissue engineering. Cultured stromal cells are a mixture of cells at different stages of commitment and expansion capability, leading to a heterogeneous cell population that each time can differ in the potential to form in vivo bone. A parameter that predicts for in vivo bone forming capacity is thus far lacking. We employed single colony-derived BMSC cultures to identify such predictive parameters. Using limiting dilution, we have produced sixteen single CFU-F derived BMSC cultures from human bone marrow and found that only five of these formed bone in vivo. The single colony-derived BMSC strains were tested for proliferation, osteogenic-, adipogenic- and chondrogenic differentiation capacity and the expression of a variety of associated markers. The only robust predictors of in vivo bone forming capacity were the induction of alkaline phosphatase, (ALP) mRNA levels and ALP activity during in vitro osteogenic differentiation. The predictive value of in vitro ALP induction was confirmed by analyzing “bulk-cultured” BMSCs from various bone marrow biopsies. Our findings show that in BMSCs, the additional increase in ALP levels over basal levels during in vitro osteogenic differentiation is predictive of in vivo performance.

Published
2014

6. Mesenchymal stem cell therapy in proteoglycan induced arthritis

Author

Infection & Immunity, Risk Assessment, dI&I RA-I&I I&I, LS Immunologie, Swart, J. F., de Roock, S., Hofhuis, F. M., Rozemuller, H., van den Broek, T., Moerer, P., Broere, F., van Wijk, F., Kuis, W., Prakken, B. J., Martens, a.c.m, Wulffraat, N. M., Infection & Immunity, Risk Assessment, dI&I RA-I&I I&I, LS Immunologie, Swart, J. F., de Roock, S., Hofhuis, F. M., Rozemuller, H., van den Broek, T., Moerer, P., Broere, F., van Wijk, F., Kuis, W., Prakken, B. J., Martens, a.c.m, and Wulffraat, N. M.

Published
2015

7. In vitro induction of alkaline phosphatase levels predicts in vivo bone forming capacity of human bone marrow stromal cells.

Author

Dep Gezondheidszorg Paard, Geneeskunde van gezelschapsdieren, CSCA TR1, Tissue Repair, Prins, H.J., Braat, A.K., Gawlitta, D., Dhert, W.J.A., Egan, D.A., Tijssen-Slump, E., Yuan, H., Coffer, P.J, Rozemuller, H., Martens, A.C.M., Dep Gezondheidszorg Paard, Geneeskunde van gezelschapsdieren, CSCA TR1, Tissue Repair, Prins, H.J., Braat, A.K., Gawlitta, D., Dhert, W.J.A., Egan, D.A., Tijssen-Slump, E., Yuan, H., Coffer, P.J, Rozemuller, H., and Martens, A.C.M.

Published
2014

8. Autologous Stem Cell Transplantation Restores Immune Tolerance in Experimental Arthritis by Renewal and Modulation of the Teff Cell Compartment

Author

Strategic Infection Biology, Dep Infectieziekten Immunologie, I&I RATIA-SIB, Delemarre, E.M., Roord, S.T.A., van den Broek, T., Zonneveld - Huijssoon, E., de Jager, W., Rozemuller, H., Martens, A.C.M., Broere, F., Wulffraat, N.M., Glant, T., Prakken, B.J., van Wijk, F., Strategic Infection Biology, Dep Infectieziekten Immunologie, I&I RATIA-SIB, Delemarre, E.M., Roord, S.T.A., van den Broek, T., Zonneveld - Huijssoon, E., de Jager, W., Rozemuller, H., Martens, A.C.M., Broere, F., Wulffraat, N.M., Glant, T., Prakken, B.J., and van Wijk, F.

Published
2014

10. Non-invasive imaging of mouse hepatitis coronavirus infection reveals determinants of viral replication and spread in vivo

Author

Raaben, M., Prins, M., Martens, A.C.M., Rottier, P.J.M., de Haan, C.A.M., Strategic Infection Biology, and Dep Infectieziekten Immunologie

Abstract

Bioluminescence imaging (BLI) is a powerful new method to study virus dissemination in the live animal. Here we used this method to monitor the spatial and temporal progression of mouse hepatitis coronavirus (MHV) infection in mice using luciferase-expressing viruses. Upon intranasal inoculation, virus replication could initially be observed in the nasal cavity and the cervical lymph nodes, after which the infection spread to the brain and frequently to the eyes. The kinetics of virus spread to and clearance from the brain appeared to depend on the inoculation dose. After intraperitoneal inoculation, virus replication was predominantly observed in the liver and occasionally in the intestines, but interestingly also in the tail and paws. BLI thus elucidated new anatomic locations of virus replication. Furthermore, MHV dissemination was shown to be critically depended on the viral spike protein, but also on the mouse strain used. Widespread dissemination was observed in mice lacking a functional type I interferon response. The importance of the type I interferon system in limiting viral spread was also demonstrated by the administration of type I interferons to mice. Our results provide new insights in coronavirus pathogenesis and demonstrate the potential of BLI to study coronavirus-host interactions in vivo.

Published
2009

16. Detection and organ distribution of herpes simplex virus thymidine kinase gene-transduced T cells to monitor alloreactivity after bone marrow transplantation in rats

Author

Kolen, S.M.H., Weijtens, M., Spronsen, A. van, Smulders, S., Weger, R. de, Witte, T.J.M. de, Dolstra, H., Hagenbeek, A., Wiel-van Kemenade, E. van de, and Martens, A.C.M.

Subjects
Immunotherapy, gene therapy and transplantation [UMCN 1.4]
Abstract

Item does not contain fulltext Introduction of the HSV-Tk suicide gene into allogeneic T cells offers the possibility to control developing host-reactive cells within the context of allogeneic bone marrow transplantation (BMT). Sensitive quantitative detection methods are a prerequisite to monitor genetically modified T cells in peripheral blood and tissues to study their involvement in graft-versus-host disease (GVHD)-induced lesions as well as their disappearance or persistence after ganciclovir (GCV)-induced suicide. We monitored the alloreactivity of HSV-Tk-transduced T cells after BMT by studying their in vivo distribution and quantity in peripheral blood and in tissues in a WAG/Rij into Brown Norway fully mismatched rat allogeneic BMT model. Genetically modified T cells were quantified in blood and tissues by fluorescence-activated cell sorting, immunohistochemical analysis, and real-time quantitative polymerase chain reaction (PCR) analysis. A significant increase in the number of allogeneic HSV-Tk(+) T cells was found in particular in spleen and lymph nodes and large numbers were found in tongue, skin, and intestines. In blood, an increase in HSV-Tk(+) T cells closely preceded clinical symptoms of GVHD. Real-time quantitative PCR proved to be a fast and accurate tool by which to quantify transduced T cells both in blood and tissues. This enables the study of the in vivo alloreactivity of retrovirus-transduced cells and the response of HSV-Tk-expressing T cells to GCV-induced suicide therapy. Furthermore, we showed the potential use to study specific cause-effect relationships in a broad range of animal and clinical studies involving genetically engineered cells.

Published
2003

17. The impact of cell source, culture methodology, culture location, and individual donors on gene expression profiles of bone marrow-derived and adipose-derived stromal cells

Author

Torensma, R., Prins, H.J., Schrama, E., Verwiel, E.T.P., Martens, A.C.M., Roelofs, H., Jansen, B.J.H., Torensma, R., Prins, H.J., Schrama, E., Verwiel, E.T.P., Martens, A.C.M., Roelofs, H., and Jansen, B.J.H.

Abstract

Item does not contain fulltext, Bone marrow (BM) stromal cells (MSCs), also known as mesenchymal stem cells, display a high degree of heterogeneity. To shed light on the causes of this heterogeneity, MSCs were collected from either human BM (n=5) or adipose tissue (AT) (n=5), and expanded using 2 different culture methods: one based on fetal calf serum, and one based on human platelet lysate. After initial expansion, MSCs were frozen, and the vials were transported to 3 different laboratories and grown for 1 passage using the same brand of culture plastic, medium, and supplements. Subsequently, the cells were harvested and assayed for their gene expression profile using the Affymetrix exon microarray platform. Based on gene expression profiles, the most discriminative feature was the anatomical harvesting site, followed by culture methodology. Remarkably, genes in the WNT pathway were expressed at higher levels in BM-derived MSCs than in AT-derived MSCs. Although differences were found between laboratories, cell culture location only slightly affects heterogeneity. Furthermore, individual donors contributed marginally to the observed differences in transcriptomes. Finally, BM-derived MSCs displayed the highest level of similarity, irrespective their culture conditions, when compared to AT-derived cells.

Published
2013

20. N-cadherin-mediated interaction with multiple myeloma cells inhibits osteoblast differentiation

Author

Groen, R.W.J., de Rooij, M.F.M., Kocemba, K.A., Reijmers, R.M., de Haan-Kramer, A., Overdijk, M.B., Aalders, L., Rozemuller, H., Martens, A.C.M., Bergsagel, P.L., Kersten, MJ, Pals, S.T., Spaargaren, M., Groen, R.W.J., de Rooij, M.F.M., Kocemba, K.A., Reijmers, R.M., de Haan-Kramer, A., Overdijk, M.B., Aalders, L., Rozemuller, H., Martens, A.C.M., Bergsagel, P.L., Kersten, MJ, Pals, S.T., and Spaargaren, M.

Published
2011

21. N-cadherin-mediated interaction with multiple myeloma cells inhibits osteoblast differentiation

Author

Other research (not in main researchprogram), Cancer, Regenerative Medicine and Stem Cells, CMM Sectie Celbiologie, CTI, Groen, R.W.J., de Rooij, M.F.M., Kocemba, K.A., Reijmers, R.M., de Haan-Kramer, A., Overdijk, M.B., Aalders, L., Rozemuller, H., Martens, A.C.M., Bergsagel, P.L., Kersten, MJ, Pals, S.T., Spaargaren, M., Other research (not in main researchprogram), Cancer, Regenerative Medicine and Stem Cells, CMM Sectie Celbiologie, CTI, Groen, R.W.J., de Rooij, M.F.M., Kocemba, K.A., Reijmers, R.M., de Haan-Kramer, A., Overdijk, M.B., Aalders, L., Rozemuller, H., Martens, A.C.M., Bergsagel, P.L., Kersten, MJ, Pals, S.T., and Spaargaren, M.

Published
2011

29. Bone-forming capacity of mesenchymal stromal cells when cultured in the presence of human platelet lysate as substitute for fetal bovine serum.

Author

Infection & Immunity, Regenerative Medicine and Stem Cells, CTI, MS Orthopaedie Algemeen, Prins, H., Rozemuller, H., Vonk - Griffioen, S.J., Verweij, V.G.M., Dhert, W.J.A., Slaper-Cortenbach, I.C.M., Martens, A.C.M., Infection & Immunity, Regenerative Medicine and Stem Cells, CTI, MS Orthopaedie Algemeen, Prins, H., Rozemuller, H., Vonk - Griffioen, S.J., Verweij, V.G.M., Dhert, W.J.A., Slaper-Cortenbach, I.C.M., and Martens, A.C.M.

Published
2009

30. In Vivo Matrix Production by Bone Marrow Stromal Cells Seeded on PLGA Scaffolds for Ligament Tissue Engineering

Author

Infection & Immunity, MS Orthopaedie Algemeen, CTI, van Eijk, F., Saris, D.B.F., Fedorovich, N.E., Kruyt, M.C., Willems, W.J., Verbout, A.J., Martens, A.C.M., Dhert, W.J.A., Creemers, L.B., Infection & Immunity, MS Orthopaedie Algemeen, CTI, van Eijk, F., Saris, D.B.F., Fedorovich, N.E., Kruyt, M.C., Willems, W.J., Verbout, A.J., Martens, A.C.M., Dhert, W.J.A., and Creemers, L.B.

Published
2009

33. Autologous bone marrow transplantation in autoimmune arthritis restores immune homeostasis through CD4+ CD25+ Foxp3+ regulatory T cells.

Author

Infection & Immunity, Cluster B, Reumatologie, Immunologie/Reumatologie, CTI, Roord, S.T.A., de Jager, W., Boon, L., Wulffraat, N.M., Martens, A.C.M., Prakken, A.B.J., van Wijk, F., Infection & Immunity, Cluster B, Reumatologie, Immunologie/Reumatologie, CTI, Roord, S.T.A., de Jager, W., Boon, L., Wulffraat, N.M., Martens, A.C.M., Prakken, A.B.J., and van Wijk, F.

Published
2008

34. Mesenchymal stem cells: a future for the treatment of arthritis?

Author

Infection & Immunity, Reumatologie, CTI, Immunologie/Reumatologie, Swart, J.F., Martens, A.C.M., Wulffraat, N.M., Infection & Immunity, Reumatologie, CTI, Immunologie/Reumatologie, Swart, J.F., Martens, A.C.M., and Wulffraat, N.M.

Published
2008

35. cAMP/PKA pathway activation in human mesenchymal stem cells in vitro results in robust bone formation in vivo.

Author

Infection & Immunity, CTI, Siddappa, R., Martens, A.C.M., Doorn, J., Leusink, A., Olivo, C., Licht, R., van Rijn, L., Gaspar, C., Fodde, R., Janssen, F., van Blitterswijk, C., de Boer, J., Infection & Immunity, CTI, Siddappa, R., Martens, A.C.M., Doorn, J., Leusink, A., Olivo, C., Licht, R., van Rijn, L., Gaspar, C., Fodde, R., Janssen, F., van Blitterswijk, C., and de Boer, J.

Published
2008

37. A bioluminescence imaging based in vivo model for preclinical testing of novel cellular immunotherapy strategies to improve the graft-versus-myeloma effect.

Author

Circulatory Health, Infection & Immunity, CTI, Poli Van Creveldkliniek Medisch, Centraal Diagnostisch Laboratorium, Rozemuller, H., van der Spek, E., Bogers - Boer, L.H., Zwart, M.C., Verweij, V.G.M., Emmelot, M.E., Groen, R.W., Spaapen, R.M., Bloem, A.C., Lokhorst, H.M., Mutis, T., Martens, A.C.M., Circulatory Health, Infection & Immunity, CTI, Poli Van Creveldkliniek Medisch, Centraal Diagnostisch Laboratorium, Rozemuller, H., van der Spek, E., Bogers - Boer, L.H., Zwart, M.C., Verweij, V.G.M., Emmelot, M.E., Groen, R.W., Spaapen, R.M., Bloem, A.C., Lokhorst, H.M., Mutis, T., and Martens, A.C.M.

Published
2008

40. Human regulatory T cells control xenogeneic graft-versus-host disease induced by autologous T cells in RAG2-/-gammac-/- immunodeficient mice.

Author

Advanced drug delivery systems/drug targeting, Dep Farmaceutische wetenschappen, Mutis, T., van Rijn, R.S., Simonetti, E.R., Aarts-Riemens, T., Emmelot, M.E., van Bloois, L., Martens, A.C.M., Verdonck, L.F., Ebeling, S.B., Advanced drug delivery systems/drug targeting, Dep Farmaceutische wetenschappen, Mutis, T., van Rijn, R.S., Simonetti, E.R., Aarts-Riemens, T., Emmelot, M.E., van Bloois, L., Martens, A.C.M., Verdonck, L.F., and Ebeling, S.B.

Published
2006

42. Acetyldinaline: A new oral cytostatic drug with impressive differential activity against leukemic cells and normal stem cells - Preclinical studies in a relevant rat model for human acute myelocytic leukemia

Author

El-Beltagi, H.M., Martens, A.C.M., Lelieveld, P., Haroun, E.A., Hagenbeek, A., and Instituut voor Toegepaste Radiobiologie en Immunologie TNO

Subjects
Male, Dose-Response Relationship, Drug, Animal, Cell Survival, Administration, Oral, Antineoplastic Agents, Bone Marrow Cells, Cell Differentiation, Leukemia, Myelocytic, Acute, Phenylenediamines, Hematopoietic Stem Cells, Drug Administration Schedule, Clone Cells, Rats, Disease Models, Animal, Bone Marrow, Rats, Inbred BN, Tumor Cells, Cultured, Drug Evaluation, Comparative Study, Drug Screening Assays, Antitumor, Support, Non-U.S. Gov't
Abstract

Acetyldinaline [CI-994; GOE 5549; PD 123 654; 4-acetylamino-N-(2'- aminophenyl)-benzamide] is the acetylated derivative form of the original compound Dinaline (GOE 1734; PD 104 208). The efficacy and toxicity of Acetyldinaline for remission-induction treatment of leukemia were evaluated and compared with those observed in previous studies of Dinaline in the Brown Norway acute myelocytic leukemia, as a preclinical model for human acute myelocytic leukemia. There were three treatment groups. Leukemic animals received either 1 or 2 courses of 5 daily p.o. administrations of Acetyldinaline with a 'full dose' of 23.7 mg/kg once daily (first group), a twice daily 'half dose' of 11.85 mg/kg with an interval of 8 h (second group), or a 'half dose' of 11.85 mg/kg once daily (third group). The drug- free interval between the 2 courses was 2 or 9 days. With repeated daily p.o. administrations of 23.7 mg/kg either in a single daily dose or a split daily dose of 2 x 11.85 mg/kg for 1 course, at least an 8-log leukemic cell kill was achieved. In contrast, with these treatment schedules, less than a 1-log cell kill of normal pluripotent hemopoietic stem cells (CFU-S) in the femoral bone marrow was found. Split daily dose treatment was more effective resulting in 37.5% cures, while no cures were observed with the single daily treatment for one course. Treatment with single daily dose of 23.7 mg/kg or a split daily dose of 2 x 11.85 mg/kg for 2 courses, with either a 2- or 9-day interval inbetween, resulted in lethal toxicity in most of rats. This result was comparable with that previously observed after equimolar doses of Dinaline (20 mg/kg). The half-dose once daily treatment with Acetyldinaline (11.85 mg/kg) for 1 or 2 cycles resulted in about a 4.5 or >8-log leukemic cell kill, respectively. Toxic side effects, i.e., damage to the gastro- intestinal tract and hemorrhages in the lungs, were more pronounced with full dose either in the single or the split daily dose regimen. No significant toxicity was observed at the half-dose treatment once daily. In conclusion, the impressive differential activity against leukemic cells and normal stem cells observed in this relevant rat model for human acute myelocytic leukemia warrants the introduction of this compound in clinical phase I/II studies. Chemicals/CAS: acetyldinaline, 112522-64-2; Antineoplastic Agents; dinaline, 58338-59-3; Phenylenediamines

Published
1993

45. The use of FISH with chromosome-specific repetitive DNA probes for the follow-up of leukemia patients

Author

Arkesteijn, G.J.A. (Ger), Erpelinck, S.L.A. (Stephen), Martens, A.C.M. (Anton), Hagemeijer, A. (Anne), Hagenbeek, A. (Anton), Arkesteijn, G.J.A. (Ger), Erpelinck, S.L.A. (Stephen), Martens, A.C.M. (Anton), Hagemeijer, A. (Anne), and Hagenbeek, A. (Anton)

Abstract

The use of fluorescence in situ hybridization (FISH) for the purpose of repeated follow-up examination of bone marrow samples from 38 leukemia patients was investigated. On the basis of conventional cytogenetic analysis, patients with acute leukemia whose leukemic cells carried numerical chromosomal aberrations were selected and followed with repetitive DNA probes that specifically hybridize to one chromosome type. Repeated cytogenetic metaphase analyses would have been laborious and not sensitive or quantitative enough to follow declining numbers of aberrant cells. FISH, as an interphase cytogenetic technique, provides a rapid and simple alternative with high sensitivity. Although FISH data before and after chemotherapy were in agreement with bone marrow cytology in 30 of 38 patients, discrepancies were noticed in specific cases. These could be explained by the presence of cytogenetically distinct subclones that behave differently during treatment, the presence of differentiated leukemic cells, changes in the chromosomal constitution caused by clonal relapse, or the fact that a numerical aberration is found by conventional chromosome banding analysis while the target region to which the probe is directed is still present in the nucleus as a diploid set.

Published
1996
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46. Chromosome specific DNA hybridization in suspension for flow cytometric detection of chimerism in bone marrow transplantation and leukemia

Author

Arkesteijn, G.J.A. (Ger), Erpelinck, S.L.A. (Stephen), Martens, A.C.M. (Anton), Hagenbeek, A. (Anton), Arkesteijn, G.J.A. (Ger), Erpelinck, S.L.A. (Stephen), Martens, A.C.M. (Anton), and Hagenbeek, A. (Anton)

Abstract

Flow cytometry was used to measure the fluorescence intensity of nuclei that were subjected to fluorescent in situ hybridization in suspension with chromosome specific DNA probes. Paraformaldehyde-fixed nuclei were protein digested with trypsin and hybridized simultaneously with a biotin-and DIG labeled chromosome specific centromere probe. A number of probes were tested in the suspension hybridizations. The method yielded fluorescent hybridization signals that allow discrimination between Y chromosome positive and negative nuclei when analyzed by flow cytometry. The method is especially suited for analysis of bone marrow cells derived from patients who have received a sex-mismatched allogeneic bone marrow transplantation. Male leukemia cells with a trisomy for chromosome 8 were mixed with normal female cells and simultaneously hybridized in suspension with a DIG labeled probe specific for chromosome 8 and the biotin labeled Y chromosome probe. Y chromosome positive or negative nuclei were so

Published
1995
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50. Genetic marking with the ΔLNGFR-gene for tracing goat cells in bone tissue engineering

Author

Kruyt, M.C., Stijns, M.M.C., Fedorovich, N.E., de Bruijn, J.D., van Blitterswijk, C.A., Verbout, A.J., Rozemuller, H., Hagenbeek, A., Dhert, W.J.A., and Martens, A.C.M.

Subjects
GENETIC markers, TISSUE engineering, STEM cells, IMMUNE system
Abstract

The use of bone marrow derived stromal cells (BMSC''s) for bone tissue engineering has gained much attention as an alternative for autologous bone grafting. Little is known however, about the survival and differentiation of the cells, especially in the clinical application. The aim of this study was to develop a method to trace goat BMSC''s in vivo. We investigated retroviral genetic marking, which allows stable expression of the label with cell division.Goat BMSC''s were subjected to an amphotropic envelope containing a MoMuLV-based vector expressing the human low affinity nerve growth factor receptor (ΔLNGFR). Labeling efficiency and effect on the cells were analyzed. Furthermore, transduced cells were seeded onto porous ceramic scaffolds, implanted subcutaneously in nude mice and examined after successive implantation periods.Flow cytometry indicated a transduction efficiency of 40–60%. Immunohistochemistry showed survival and subsequent bone formation of the gene-marked cells in vivo. Besides, marked cells were also found in cartilage and fibrous tissue. These findings indicate the maintenance of the precursor phenotype following gene transfer as well as the ability of the gene to be expressed following differentiation. We conclude that retroviral gene marking with ΔLNGFR is applicable to trace goat BMSC''s in bone tissue engineering research. [Copyright &y& Elsevier]

Published
2004
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