Human embryonic stem cells (hESCs) have two properties of interest for the development of cell therapies: self-renewal and the potential to differentiate into all major lineages of somatic cells in the human body. Widespread clinical application of hESC-derived cells will require culture methods that are low-cost, robust, scalable and use chemically defined raw materials. Here we describe synthetic peptide-acrylate surfaces (PAS) that support self-renewal of hESCs in chemically defined, xeno-free medium. H1 and H7 hESCs were successfully maintained on PAS for over ten passages. Cell morphology and phenotypic marker expression were similar for cells cultured on PAS or Matrigel. Cells on PAS retained normal karyotype and pluripotency and were able to differentiate to functional cardiomyocytes on PAS. Finally, PAS were scaled up to large culture-vessel formats. Synthetic, xeno-free, scalable surfaces that support the self-renewal and differentiation of hESCs will be useful for both research purposes and development of cell therapies.
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We would like to thank our sponsors from Corning Incorporated, J. Mooney and M. McFarland, and from Geron Corporation, J. Lebkowski and A.H. Davies. From Corning Incorporated we thank A. Frutos for critical reading and helpful comments on the manuscript, M. Lewis for his chemistry expertise, T. Garvey for his assistance in PAS fabrication, P. Gagnon and P. Szlosek for substrate plate supply and T. Heck for his confocal imaging help. From Geron Corporation we thank K. Delavan-Boorsma and S. Edell for their support of in vivo studies.
Z.M., J.L.W., P.D.-S., J.P.B., S.P., D.M.W., A.G.F., Y.Z., C.S., A.M., J.N. and P.W. are employees of Corning Incorporated. J.Y., L.Q., C.A.P. and R.B. are employees of Geron Corporation.
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Melkoumian, Z., Weber, J., Weber, D. et al. Synthetic peptide-acrylate surfaces for long-term self-renewal and cardiomyocyte differentiation of human embryonic stem cells. Nat Biotechnol 28, 606–610 (2010) doi:10.1038/nbt.1629
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