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

被引:72
|
作者
Sugii, Shigeki [1 ,2 ]
Kida, Yasuyuki [2 ]
Berggren, W. Travis
Evans, Ronald M. [1 ,2 ]
机构
[1] Salk Inst Biol Studies, Howard Hughes Med Inst, La Jolla, CA 92037 USA
[2] Salk Inst Biol Studies, Gene Express Lab, La Jolla, CA 92037 USA
基金
美国国家卫生研究院;
关键词
HUMAN SOMATIC-CELLS; HUMAN FIBROBLASTS; GENE DELIVERY; GENERATION; INDUCTION; VECTOR; SYSTEM; TISSUE; FAT; DIFFERENTIATION;
D O I
10.1038/nprot.2010.199
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:346 / 358
页数:13
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