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Chemically defined conditions for human iPSC derivation and culture

Authors :
Mitchell D. Probasco
Daniel R. Gulbranson
Kimberly Smuga-Otto
Jessica Antosiewicz-Bourget
Zhonggang Hou
Guokai Chen
Victor Ruotti
Jennifer M. Bolin
Sara E. Howden
James A. Thomson
Ryan J. Wagner
Garrett O. Lee
Nicholas E. Propson
Nicole R. Diol
Joyce M.C. Teng
Source :
Nature methods
Publication Year :
2011
Publisher :
Springer Science and Business Media LLC, 2011.

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.

Details

ISSN :
15487105 and 15487091
Volume :
8
Database :
OpenAIRE
Journal :
Nature Methods
Accession number :
edsair.doi.dedup.....537404210d8fb638ada6d99acf07b1aa
Full Text :
https://doi.org/10.1038/nmeth.1593