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


ABSTRACT: 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 culture-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 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.

SUBMITTER: Fluri DA 

PROVIDER: S-EPMC4954777 | biostudies-literature | 2012 Mar

REPOSITORIES: biostudies-literature

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

Fluri David A DA   Tonge Peter D PD   Song Hannah H   Baptista Ricardo P RP   Shakiba Nika N   Shukla Shreya S   Clarke Geoffrey G   Nagy Andras A   Zandstra Peter W PW  

Nature methods 20120325 5


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 culture-reprogrammed iPSCs (SiPSCs) could be differentiated into all three germ layers in vitro and contributed to chimeric embryos in vivo. SiPSC generation  ...[more]

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