Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture

Hui Hsuan Kuo; Xiaozhi Gao; Jean Marc DeKeyser; K. Ashley Fetterman; Emily A. Pinheiro; Carly J. Weddle; Hananeh Fonoudi; Michael V. Orman; Marisol Romero-Tejeda; Mariam Jouni; Malorie Blancard; Tarek Magdy; Conrad L. Epting; Alfred L. George; Paul W. Burridge

doi:10.1016/j.stemcr.2019.12.007

Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture

Hui Hsuan Kuo, Xiaozhi Gao, Jean Marc DeKeyser, K. Ashley Fetterman, Emily A. Pinheiro, Carly J. Weddle, Hananeh Fonoudi, Michael V. Orman, Marisol Romero-Tejeda, Mariam Jouni, Malorie Blancard, Tarek Magdy, Conrad L. Epting, Alfred L. George, Paul W. Burridge^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

95 Scopus citations

Abstract

Human induced pluripotent stem cell (hiPSC) culture has become routine, yet the cost of pluripotent cell media, frequent medium changes, and the reproducibility of differentiation have remained restrictive. Here, we describe the formulation of a hiPSC culture medium (B8) as a result of the exhaustive optimization of medium constituents and concentrations, establishing the necessity and relative contributions of each component to the pluripotent state and cell proliferation. The reagents in B8 represent only 3% of the costs of commercial media, made possible primarily by the in-lab generation of three E. coli-expressed, codon-optimized recombinant proteins: fibroblast growth factor 2, transforming growth factor β3, and neuregulin 1. We demonstrate the derivation and culture of 34 hiPSC lines in B8 as well as the maintenance of pluripotency long term (over 100 passages). This formula also allows a weekend-free feeding schedule without sacrificing capacity for differentiation.

Original language	English (US)
Pages (from-to)	256-270
Number of pages	15
Journal	Stem cell reports
Volume	14
Issue number	2
DOIs	https://doi.org/10.1016/j.stemcr.2019.12.007
State	Published - Feb 11 2020

Funding

This work was supported by NIH NCI grant R01 CA220002 , American Heart Association Transformational Project Award 18TPA34230105 , a Dixon Foundation Translational Research Grants Innovation Award (P.W.B.), and the Fondation Leducq (P.W.B., A.L.G.).

Keywords

FGF2
chemically defined
culture media
differentiation
human induced pluripotent stem cell
pluripotent state
weekend-free

ASJC Scopus subject areas

Biochemistry
Genetics
Developmental Biology
Cell Biology

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10.1016/j.stemcr.2019.12.007

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Kuo, H. H., Gao, X., DeKeyser, J. M., Fetterman, K. A., Pinheiro, E. A., Weddle, C. J., Fonoudi, H., Orman, M. V., Romero-Tejeda, M., Jouni, M., Blancard, M., Magdy, T., Epting, C. L., George, A. L., & Burridge, P. W. (2020). Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture. Stem cell reports, 14(2), 256-270. https://doi.org/10.1016/j.stemcr.2019.12.007

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Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture

Abstract

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Keywords

ASJC Scopus subject areas

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