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A simple tool to improve pluripotent stem cell differentiation

Nature Methods volume 10, pages 553–556 (2013)Cite this article

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Abstract

We describe a method to help overcome restrictions on the differentiation propensities of human pluripotent stem cells. Culturing pluripotent stem cells in dimethylsulfoxide (DMSO) activates the retinoblastoma protein, increases the proportion of cells in the early G1 phase of the cell cycle and, in more than 25 embryonic and induced pluripotent stem cell lines, improves directed differentiation into multiple lineages. DMSO treatment also improves differentiation into terminal cell types in several cell lines.

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Figure 1: DMSO treatment enhances differentiation in hESC and hiPSC lines.
Figure 2: DMSO treatment improves differentiation into all germ layers across multiple hESC and hiPSC lines.
Figure 3: DMSO-treated cells have activated retinoblastoma protein (Rb) and an increased proportion of early G1-stage cells.

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Acknowledgements

We thank E. Scadden for her excellent technical support; R. Maehr, P. Makhijani and J. Millman of Harvard University for providing many of the hiPSC lines; K. Lam for assistance with the Cellomics high-content screening system; and A. Meissner, K. Eggan, D. Cohen, R. Maehr, J. Rivera-Feliciano, B. Blum, J. Sneddon and Q. Peterson for their comments on the manuscript. This work was supported by the Harvard Stem Cell Institute, the Howard Hughes Medical Institute, the Leona M. and Harry B. Helmsley Charitable Trust, NovoNordisk and the US National Institutes of Health (1R24DK092758 and 1U01HL10040804).

Author information

Authors and Affiliations

  1. Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA

    Sundari Chetty, Felicia Walton Pagliuca, Christian Honore & Douglas A Melton

  2. CureMeta, Boston, Massachusetts, USA

    Anastasie Kweudjeu

  3. BetaLogics Venture, Janssen R&D LLC, Raritan, New Jersey, USA

    Alireza Rezania

  4. Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA

    Douglas A Melton

Authors
  1. Sundari Chetty
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  2. Felicia Walton Pagliuca
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  3. Christian Honore
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  4. Anastasie Kweudjeu
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  5. Alireza Rezania
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  6. Douglas A Melton
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Contributions

S.C. and D.A.M. conceived and designed the research, analyzed the data and wrote the manuscript. S.C., F.W.P., C.H., A.K. and A.R. performed the experiments.

Corresponding author

Correspondence to Douglas A Melton.

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Competing interests

A.R. is an employee of BetaLogics, a division of Janssen Research & Development that is developing commercial products based upon pancreatic differentiation of human embryonic stem cells.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–12, Supplementary Tables 1 and 2 and Supplementary Note (PDF 2913 kb)

DMSO treatment of hESCs generates functional cardiomyocytes (video 1 of 4)

DMSO treatment of the HUES6 cell line enhances the potential for cardiomyocyte differentiation and induces functionality by promoting contractile properties. (MOV 2020 kb)

DMSO treatment of hESCs generates functional cardiomyocytes (video 2 of 4)

DMSO treatment of the HUES6 cell line enhances the potential for cardiomyocyte differentiation and induces functionality by promoting contractile properties. (MOV 13785 kb)

DMSO treatment of hESCs generates functional cardiomyocytes (video 3 of 4)

DMSO treatment of the HUES6 cell line enhances the potential for cardiomyocyte differentiation and induces functionality by promoting contractile properties. (MOV 6905 kb)

DMSO treatment of hESCs generates functional cardiomyocytes (video 4 of 4)

DMSO treatment of the HUES6 cell line enhances the potential for cardiomyocyte differentiation and induces functionality by promoting contractile properties. (MOV 5436 kb)

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Chetty, S., Pagliuca, F., Honore, C. et al. A simple tool to improve pluripotent stem cell differentiation. Nat Methods 10, 553–556 (2013). https://doi.org/10.1038/nmeth.2442

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