Derivation, expansion and differentiation of induced pluripotent stem cells in continuous suspension cultures

作者：Fluri David A; Tonge Peter D; Song Hannah; Baptista Ricardo P; Shakiba Nika; Shukla Shreya; Clarke Geoffrey; Nagy Andras; Zandstra Peter W^*

来源：Nature Methods, 2012, 9(5): 509-U145.

DOI：10.1038/NMETH.1939

We describe 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 culturere-programmed 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 culture. Seamless integration of SiPSC reprogramming and directed differentiation enabled scalable production of beating cardiac cells in a continuous single cell- and small aggregate-based process. This method is an important step toward the development of robust PSC generation, expansion and differentiation technology.

• 出版日期2012-5