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Feeder-dependent and feeder-independent iPS cell derivation from human and mouse adipose stem cells

Nature Protocols volume 6, pages 346–358 (2011)Cite this article

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Abstract

Adipose tissue is an abundantly available source of proliferative and multipotent mesenchymal stem cells with promising potential for regenerative therapeutics. We previously demonstrated that both human and mouse adipose-derived stem cells (ASCs) can be reprogrammed into induced pluripotent stem cells (iPSCs) with efficiencies higher than those that have been reported for other cell types. The ASC-derived iPSCs can be generated in a feeder-independent manner, representing a unique model to study reprogramming and an important step toward establishing a safe, clinical grade of cells for therapeutic use. In this study, we provide a detailed protocol for isolation, preparation and transformation of ASCs from fat tissue into mouse iPSCs in feeder-free conditions and human iPSCs using feeder-dependent or feeder/xenobiotic-free processes. This protocol also describes how ASCs can be used as feeder cells for maintenance of other pluripotent stem cells. ASC derivation is rapid and can be completed in <1 week, with mouse and human iPS reprogramming times averaging 1.5 and 2.5 weeks, respectively.

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Figure 1: Morphology of ASCs in culture observed with ×40 magnification on an inverted phase-contrast microscope.
Figure 2: Derivation of mouse ASC-derived iPSCs.
Figure 3: Derivation of hASC-derived iPSCs.

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Acknowledgements

We thank J.M. Gimble and members of the Izpisúa Belmonte laboratory for helpful discussions, L. Ong and S. Ganley for administrative assistance and R. Yu for advice on and editing of the manuscript. This work was supported by grants from the National Institutes of Health (HD027183, DK057978 and DK062434), California Institute for Regenerative Medicine (RB2-01530) and Howard Hughes Medical Institute.

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Author notes

  1. Shigeki Sugii & W Travis Berggren

    Present address: Present address: Singapore Bioimaging Consortium and Duke–NUS (National University of Singapore) Graduate Medical School, Singapore.,

  2. Shigeki Sugii and Yasuyuki Kida: These authors contributed equally to this work.

Authors and Affiliations

  1. Howard Hughes Medical Institute, The Salk Institute for Biological Studies, La Jolla, California, USA

    Shigeki Sugii & Ronald M Evans

  2. Gene Expression Laboratory, The Salk Institute for Biological Studies, La Jolla, California, USA

    Shigeki Sugii, Yasuyuki Kida & Ronald M Evans

  3. Stem Cell Core, The Salk Institute for Biological Studies, La Jolla, California, USA

    Shigeki Sugii

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Contributions

S.S. and Y.K. designed and performed the experimental procedures. S.S., Y.K. and W.T.B. wrote the protocol. R.M.E. was the project leader, obtained funding, and reviewed and edited the protocol.

Corresponding author

Correspondence to Ronald M Evans.

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The authors have filed a patent application on some of the methods described in this article.

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Sugii, S., Kida, Y., Berggren, W. et al. Feeder-dependent and feeder-independent iPS cell derivation from human and mouse adipose stem cells. Nat Protoc 6, 346–358 (2011). https://doi.org/10.1038/nprot.2010.199

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