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Abstract

Human induced pluripotent stem cells (iPSCs) present exciting opportunities for studying development and for in vitro disease modeling. However, reported variability in the behavior of iPSCs has called their utility into question. We established a test set of 16 iPSC lines from seven individuals of varying age, sex and health status, and extensively characterized the lines with respect to pluripotency and the ability to terminally differentiate. Under standardized procedures in two independent laboratories, 13 of the iPSC lines gave rise to functional motor neurons with a range of efficiencies similar to that of human embryonic stem cells (ESCs). Although three iPSC lines were resistant to neural differentiation, early neuralization rescued their performance. Therefore, all 16 iPSC lines passed a stringent test of differentiation capacity despite variations in karyotype and in the expression of early pluripotency markers and transgenes. This iPSC and ESC test set is a robust resource for those interested in the basic biology of stem cells and their applications.

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Change history

  • 11 February 2011

    In the version of this article initially published online, a funder was inadvertently omitted from the Acknowledgments. The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank H. Mitsumoto, J. Montes, P. Kaufmann and J. Andrews for collecting skin biopsies; K. Koszka, A. Sproul, A. Hon and A. Garcia-Diaz for technical assistance; M. Park, A. Meissner and C. Bock for manuscript assistance, as well as S. Brenner-Morton and T. Jessell for providing Islet antibodies. This work was funded by Project A.L.S., P2ALS, NYSTEM and the National Institutes of Health (NIH) GO grant 1RC2 NS069395-01. G.L.B. is a Harvard Stem Cell Institute/NIH Trainee. E.K. is an EMBO Postdoctoral Fellow. B.J.W. is supported by NIH Training Grant 5T32GM007592. C.J.W. is supported by grants from the National Institute of Neurological Disorders and Stroke and the National Institute of Child Health and Development. K.E. is a Howard Hughes Medical Institute early career scientist.

Author information

Author notes

    • John T Dimos

    Present address: iPierian, Inc., South San Francisco, California, USA.

    • Gabriella L Boulting
    • , Evangelos Kiskinis
    • , Gist F Croft
    • , Mackenzie W Amoroso
    •  & Derek H Oakley

    These authors contributed equally to this work.

Affiliations

  1. The Howard Hughes Medical Institute, Cambridge, Massachusetts, USA.

    • Gabriella L Boulting
    • , Evangelos Kiskinis
    • , Mariko Yamaki
    •  & Kevin Eggan
  2. Harvard Stem Cell Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA.

    • Gabriella L Boulting
    • , Evangelos Kiskinis
    • , Mariko Yamaki
    • , Lance Davidow
    • , Shravani Mikkilineni
    •  & Kevin Eggan
  3. Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.

    • Gabriella L Boulting
    • , Christopher T Rodolfa
    • , John T Dimos
    • , Shravani Mikkilineni
    •  & Kevin Eggan
  4. Project A.L.S./Jenifer Estess Laboratory for Stem Cell Research, Columbia University, New York, New York, USA.

    • Gist F Croft
    • , Mackenzie W Amoroso
    • , Derek H Oakley
    • , Christopher E Henderson
    •  & Hynek Wichterle
  5. Departments of Pathology, Neurology and Neuroscience, Columbia University, Center for Motor Neuron Biology and Disease (MNC), and Columbia Stem Cell Initiative (CSCI), New York, New York, USA.

    • Gist F Croft
    • , Mackenzie W Amoroso
    • , Derek H Oakley
    • , Christopher E Henderson
    •  & Hynek Wichterle
  6. Program in Neurobiology and FM Kirby Neurobiology Center, Children's Hospital Boston, Boston, Massachusetts, USA.

    • Brian J Wainger
    •  & Clifford J Woolf
  7. Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.

    • Brian J Wainger
    •  & Clifford J Woolf
  8. Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.

    • Brian J Wainger
  9. Departments of Physiology and Cellular Biophysics, and Neuroscience, Columbia University, New York, New York, USA.

    • Damian J Williams
    •  & Amy B MacDermott
  10. The New York Stem Cell Foundation, Inc. (NYSCF), New York, New York, USA.

    • David J Kahler

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Contributions

G.F.C., M.W.A. and D.H.O. maintained human fibroblasts. C.T.R. and J.T.D. reprogrammed all iPSC lines. G.L.B. and E.K. expanded all iPSC lines. G.L.B. and E.K. led and contributed equally to all other experiments and analyses in the Eggan laboratory. G.F.C., M.W.A. and D.H.O. led and contributed equally to all other experiments and analyses in the Project ALS laboratory. D.J.K. did FC analysis. A.B.M., D.J.W. and D.H.O. designed and carried out Ca2+ imaging. B.J.W., G.L.B. and C.J.W. did recordings. M.Y. assisted with teratomas. L.D. assisted with quantitative analysis. S.M. assisted with stem cell culture. G.L.B., E.K., K.E., G.F.C., M.W.A., D.H.O., C.E.H. and H.W. conceived the experiments and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Hynek Wichterle or Kevin Eggan.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Tables 1–7, Supplementary Figs. 1–8 and Supplementary Methods

Videos

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    Supplementary Video 1

    Spontaneous activity in iPS 18c-derived motor neurons.

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DOI

https://doi.org/10.1038/nbt.1783