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A new expansion system for induced pluripotent stem cells in stirred suspension bioreactors
Author(s): 
Pages: 707-709
Year: Issue:  4
Journal: Chinese Journal of Experimental Surgery

Keyword:  Pluripotent stem cellsCultureCell expansion;
Abstract: Objective To establish protocols for expanding human induced pluripotent stem cells (iPSCs) as undifferentiated aggregates in stirred suspension bioreactors and illustrate that such protocols can be adapted for long term culture of human iPSCs.Methods The human iPSCs lines maintained on the Matrigel-coated plates were collected,seeded in stirred suspension bioreactor,and cultured for 8 passages,4 days each passage.Cell number and viability were calculated and the immunoflurescence analysis and gene expression profiles of pluripotency markers including Oct4,Nanog,sex determining region Y-box 2 (Sox2) and alkaline phosphatase (ALP) were conducted at the start and the end of the experiment.In vitro differentiation into hepatocytes was performed at the end of the long term culture.Results The doubling time of iPSCs in stirred suspension bioreactors was (20.82 ± 1.27) h,and the cells formed cell aggregates whose diameter got (184.1 ± 7.2) μm after suspension culturing for 4 days.Stirred suspension bioreactors yielded 1 × 1014-fold expansion of undifferentiated pluripotent iPSCs over culture of 32 days.The iPSCs still expressed the pluripotency markers over extended culture time in suspension.In vitro differentiation into hepatocytes,further confirmed the functional and undifferentiated pluripotent iPSC aggregates following long-term passaging.Conclusion Stirred suspension bioreactor system represents an efficient process for the large-scale expansion of highly pluripotent human iPSCs populations,which is an important and the first step in their clinical application.
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