Derivation, expansion and differentiation of induced pluripotent stem cells in continuous suspension cultures.
Derivation, expansion and differentiation of induced pluripotent stem cells in continuous suspension cultures.
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DOI：
10.1038/nmeth.1939
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发表时间：
2012-03-25
期刊：
影响因子：
48
通讯作者：
Zandstra PW
中科院分区：
文献类型：
--
作者：
Fluri DA;Tonge PD;Song H;Baptista RP;Shakiba N;Shukla S;Clarke G;Nagy A;Zandstra PW
我们证明了在无血清和无饲养层的搅拌悬浮培养物中从终末分化的小鼠细胞衍生出诱导多能干细胞（iPSC）。全局基因表达的时间分析揭示了悬浮中重编程的细胞和粘附依赖性条件下重编程的细胞之间的高度相关性。悬浮（S）重编程的 iPSC（SiPSC）可以在体外分化为所有三个胚层，并有助于在体内形成嵌合胚胎。 SiPSC 的产生允许根据重编程因子表达细胞在悬浮液中的差异存活和增殖来有效选择它们。 SiPSC 重编程和定向分化的无缝集成使得能够在连续的单细胞和小聚集过程中大规模生产功能和表型明确的心肌细胞。该方法是开发强大的 PSC 生成、扩展和分化技术的重要一步。
We demonstrate derivation of induced pluripotent stem cells (iPSCs) from terminally differentiated mouse cells in serum- and feeder-free stirred suspension cultures. Temporal analysis of global gene expression revealed high correlations between cells reprogrammed in suspension and cells reprogrammed in adhesion-dependent conditions. Suspension (S) reprogrammed iPSCs (SiPSCs) could be differentiated into all three germ layers in vitro and contributed to chimeric embryos in vivo. SiPSC generation allowed for efficient selection of reprogramming factor expressing cells based on their differential survival and proliferation in suspension. Seamless integration of SiPSC reprogramming and directed differentiation enabled the scalable production of functionally and phenotypically defined cardiac cells in a continuous single cell- and small aggregate-based process. This method is an important step towards the development of a robust PSC generation, expansion and differentiation technology.