Synthetic polymer coatings for long-term growth of human embryonic stem cells

Abstract

We report a fully defined synthetic polymer coating, poly[2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl)ammonium hydroxide] (PMEDSAH), which sustains long-term human embryonic stem (hES) cell growth in several different culture media, including commercially available defined media. The development of a standardized, controllable and sustainable culture matrix for hES cells is an essential step in elucidating mechanisms that control hES cell behavior and in optimizing conditions for biomedical applications of hES cells.

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Figure 1: Long-term culture of H9 hES cells on PMEDSAH with MEF-conditioned media.
Figure 2: PMEDSAH supports culture of hES cells in defined medium.

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Acknowledgements

H.N. acknowledges funding from the University of Michigan Rackham Predoctoral Fellowship. J.L. gratefully acknowledges support from the National Science Foundation (NSF) in the form of a CAREER grant and funding from the NSF under the MRI program. We would like to thank J.H. Elisseeff and N. Hwang for their insightful comments throughout this study. We further thank J. Garcia-Perez, M. Morell, L.S.D. Emmett, M. Dzaman, M. Bormann, C. Pacut, N.Leff and J. MacDonald for their valuable assistance. We would also like to thank C. Smith, M. Yoshida and S. Brown for their invaluable comments on the manuscript. This research was supported by US National Institutes of Health grants P20 GM-069985, R01 DE016530 and the National Institute of Dental and Craniofacial Research T32 Tissue Engineering and Regeneration Training Program DE 07057.

Author information

L.G.V.-D. and H.N. contributed equally to this work and were involved in experimental design, performing hES cell culture experiments, data analysis and manuscript preparation. L.G.V.-D. carried out cell analysis experiments, and H.N. fabricated the polymer coatings and performed surface analysis. J.D. was involved in immunocytochemistry and RT-PCR, while N.C.N.-d.-S. conducted microarray analysis and qPCR. P.H.K. participated in manuscript preparation. K.S.O. participated in manuscript preparation and performed teratoma assays. J.L. and G.D.S. were involved in conceptual and experimental design, as well as in manuscript preparation.

Correspondence to Joerg Lahann or Gary D Smith.

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The authors declare no competing financial interests.

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Supplementary Text and Figures

Supplementary Tables 1,2, Supplementary Figs. 1–3 and Supplementary Methods (PDF 408 kb)

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Villa-Diaz, L., Nandivada, H., Ding, J. et al. Synthetic polymer coatings for long-term growth of human embryonic stem cells. Nat Biotechnol 28, 581–583 (2010) doi:10.1038/nbt.1631

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