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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Acknowledgements
This work was supported by The Charlotte Geyer Foundation, the Morgridge Institute for Research, US National Institutes of Health grant UO1ES017166 (to J.A.T.) and National Institutes of Health contract RR-05-19 (to J.A.T.). We thank K. Eastman for editorial assistance, and C. Dewey, R. Stewart and A. Elwell for their assistance with gene expression analysis.
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G.C. and J.A.T. conceived the experiment; G.C., D.R.G., J.M.B., K.S.-O. and S.E.H. performed the reprogramming; Z.H., G.C. and N.E.P. produced vitronectin; G.C., D.R.G., J.M.B., N.R.D., G.O.L. and J.A.-B. performed the cell culture test; G.C., M.D.P. and R.W. derived fibroblasts; J.M.C.T. obtained the skin biopsy; V.R. and G.C. analyzed global expression; and G.C. and J.A.T. wrote the paper.
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J.A.T. is a founder, stockowner, consultant and board member of Cellular Dynamics International (CDI), and serves as scientific advisor to and has financial interests in Tactics II Stem Cell Ventures.
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Chen, G., Gulbranson, D., Hou, Z. et al. Chemically defined conditions for human iPSC derivation and culture. Nat Methods 8, 424–429 (2011). https://doi.org/10.1038/nmeth.1593
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DOI: https://doi.org/10.1038/nmeth.1593
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