Abstract
Mesenchymal stem cells (MSCs) have been isolated from many tissues including differentiating human embryonic stem cells (hESCs). Derivation of MSCs from hESCs consists of two major steps: differentiation and isolation. In our hands, differentiation of hESCs towards MSC-enriched culture can be induced by trypsinizing hESCs into single cells and plating them on gelatin-coated plates in a culture condition that enhances survival of hESC-derived MSCs and not hESCs. The trypsinized hESCs were grown with feeder support and the medium was supplemented with basic fibroblast growth factor (FGF2) and platelet-derived growth factor (PDGF)-AB. A highly enriched MSC culture could be obtained by repeated passaging by trypsinization. The enriched MSC cultures could be further purified by limiting dilution or FACS sorting for CD105+ or CD73+ and CD24−.
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References
Wagner, W., and Ho, A.D. (2007) Mesenchymal stem cell preparations - comparing apples and oranges. Stem Cell Rev. 3, 239–248.
Brooke, G., Cook, M., Blair, C., Han, R., Heazlewood, C., Jones, B., et al. (2007) Therapeutic applications of mesenchymal stromal cells. Semin. Cell. Dev. Biol. 18, 846–858.
Dominici, M., Le Blanc, K., Mueller, I., Slaper-Cortenbach, I., Marini, F., Krause, D., et al. (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8, 315–317.
Barberi, T., Willis, L.M., Socci, N.D., and Studer, L. (2005) Derivation of multipotent mesenchymal precursors from human embryonic stem cells. PLoS Med. 2, e161.
Lian, Q., Lye, E., Suan Yeo, K., Khia Way Tan, E., Salto-Tellez, M., Liu, T.M., et al. (2007) Derivation of clinically compliant MSCs from CD105+, CD24− differentiated human ESCs. Stem Cells 25, 425–436.
Olivier, E.N., Rybicki, A.C., and Bouhassira, E.E. (2006) Differentiation of human embryonic stem cells into bipotent mesenchymal stem cells. Stem Cells 24, 1914–1922.
Caplan, A.I., and Dennis, J.E. (2006) Mesenchymal stem cells as trophic mediators. J. Cell. Biochem. 98, 1076–1084.
Sze, S.K., de Kleijn, D.P., Lai, R.C., Khia Way Tan, E., Zhao, H., Yeo, K.S., et al. (2007) Elucidating the secretion proteome of human embryonic stem cell-derived mesenchymal stem cells. Mol. Cell. Proteomics 6, 1680–1689.
Timmers, L., Lim, S.K., Arslan, F., Armstrong, J.S., Hoefer, I.E., Doevendans, P.A., et al. (2007) Reduction of myocardial infarct size by human mesenchymal stem cell conditioned medium. Stem Cell Res. 1, 129–137.
Choo, A., Ngo, A.S., Ding, V., Oh, S., and Kiang, L.S. (2008) Autogeneic feeders for the culture of undifferentiated human embryonic stem cells in feeder and feeder-free conditions. Methods Cell Biol. 86, 15–28.
Cowan, C.A., Klimanskaya, I., McMahon, J., Atienza, J., Witmyer, J., Zucker, J.P., et al. (2004) Derivation of embryonic stem-cell lines from human blastocysts. N. Engl. J. Med. 350, 1353–1356.
Thomson, J.A., Itskovitz-Eldor, J., Shapiro, S.S., Waknitz, M.A., Swiergiel, J.J., Marshall, V.S., et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.
Acknowledgments
This work was supported by funding from A*STAR. We thank members of our laboratories for their contributions to this work.
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Choo, A., Lim, S.K. (2011). Derivation of Mesenchymal Stem Cells from Human Embryonic Stem Cells. In: Nieden, N. (eds) Embryonic Stem Cell Therapy for Osteo-Degenerative Diseases. Methods in Molecular Biology, vol 690. Humana Press. https://doi.org/10.1007/978-1-60761-962-8_12
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DOI: https://doi.org/10.1007/978-1-60761-962-8_12
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