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 journalArticle

1 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 languageEnglish (US)
Pages (from-to)256-270
Number of pages15
JournalStem cell reports
Volume14
Issue number2
DOIs
StatePublished - Feb 11 2020

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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    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