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

Feeder-dependent and feeder-independent iPS cell derivation from human and mouse adipose stem cells.
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DOI:
10.1038/nprot.2010.199
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发表时间:
2011-03
期刊:
影响因子:
14.8
通讯作者:
Evans, Ronald M.
Evans, Ronald M.
中科院分区:
生物学1区
文献类型:
--
作者:
Sugii, Shigeki;Kida, Yasuyuki;Berggren, W. Travis;Evans, Ronald M.

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脂肪组织是一个丰富的来源,增殖和多能间充质干细胞与再生治疗的特殊潜力。我们先前证明,人和小鼠脂肪源性干细胞(ADS)都可以重编程为诱导多能干细胞(iPS),其效率高于其他细胞类型。ADS衍生的iPS细胞可以以不依赖饲养层的方式产生,代表了研究重编程的独特模型,也是建立安全的临床移植细胞等级的重要一步。在这里,我们提供了一个详细的协议,分离,制备和转化ADS细胞从脂肪组织转化为iPS细胞在饲养和外源性的过程。该方案还描述了如何将ADS细胞用作饲养细胞以维持其他多能干细胞。ADS衍生是快速的,可以在不到1周内完成,小鼠和人iPS重编程平均分别为1.5周和2.5周。
Adipose tissue is an abundantly available source of proliferative and multipotent mesenchymal stem cells with exceptional potential for regenerative therapeutics. We previously demonstrated that both human and mouse adipose-derived stem (ADS) cells can be reprogrammed into induced pluripotent stem (iPS) cells with efficiencies higher than have been reported for other cell types. The ADS-derived iPS cells can be generated in a feeder-independent manner, representing a unique model to study reprogramming and a significant step toward establishing a safe, clinical grade of cells for transplantation. Here we provide a detailed protocol for isolation, preparation and transformation of ADS cells from fat tissue into iPS cells in a feeder- and xenobiotic-free process. This protocol also describes how ADS cells can be used as feeder cells for maintenance of other pluripotent stem cells. ADS derivation is rapid and can be completed in less than 1 week, with mouse and human iPS reprogramming averaging 1.5 and 2.5 weeks, respectively.
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