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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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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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.
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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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DOI: https://doi.org/10.1038/nprot.2010.199
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