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Chemically defined conditions for human iPSC derivation and culture
Authors:Chen Guokai  Gulbranson Daniel R  Hou Zhonggang  Bolin Jennifer M  Ruotti Victor  Probasco Mitchell D  Smuga-Otto Kimberly  Howden Sara E  Diol Nicole R  Propson Nicholas E  Wagner Ryan  Lee Garrett O  Antosiewicz-Bourget Jessica  Teng Joyce M C  Thomson James A
Institution:Morgridge Institute for Research, Madison, Wisconsin, USA.
Abstract:We re-examine the individual components for human embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) culture and formulate a cell culture system in which all protein reagents for liquid media, attachment surfaces and splitting are chemically defined. A major improvement is the lack of a serum albumin component, as variations in either animal- or human-sourced albumin batches have previously plagued human ESC and iPSC culture with inconsistencies. Using this new medium (E8) and vitronectin-coated surfaces, we demonstrate improved derivation efficiencies of vector-free human iPSCs with an episomal approach. This simplified E8 medium should facilitate both the research use and clinical applications of human ESCs and iPSCs and their derivatives, and should be applicable to other reprogramming methods.
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