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Nat Protoc. 2011 Mar;6(3):346-58. doi: 10.1038/nprot.2010.199. Epub 2011 Feb 24.

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

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

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.

PMID:
21372815
PMCID:
PMC3089977
DOI:
10.1038/nprot.2010.199
[Indexed for MEDLINE]
Free PMC Article
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