2. Academic Validation
  3. Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture

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

  • Stem Cell Reports. 2020 Feb 11;14(2):256-270. doi: 10.1016/j.stemcr.2019.12.007.
Hui-Hsuan Kuo 1,  Xiaozhi Gao 1,  Jean-Marc DeKeyser 1,  K Ashley Fetterman 1,  Emily A Pinheiro 1,  Carly J Weddle 1,  Hananeh Fonoudi 1,  Michael V Orman 1,  Marisol Romero-Tejeda 1,  Mariam Jouni 1,  Malorie Blancard 1,  Tarek Magdy 1,  Conrad L Epting 2,  Alfred L George Jr 1,  Paul W Burridge 3
Affiliations

Affiliations

  • 1 Department of Pharmacology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA; Center for Pharmacogenomics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
  • 2 Departments of Pediatrics and Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
  • 3 Department of Pharmacology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA; Center for Pharmacogenomics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Electronic address: [email protected].
PMID: 31928950 DOI: 10.1016/j.stemcr.2019.12.007
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.

Keywords

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

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