Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture

Pijuan-Galit�, Sara; Tamm, Christoffer; Schuster, Jens; Sobol, Maria; Forsberg, Lars; Merry, Catherine L.R.; Anner�n, Cecilia

Abstract

Reliable, scalable and time-efficient culture methods are required to fully realize the clinical and industrial applications of human pluripotent stem (hPS) cells. Here we present a completely defined, xeno-free medium that supports long-term propagation of hPS cells on uncoated tissue culture plastic. The medium consists of the Essential 8 (E8) formulation supplemented with inter-?-inhibitor (I?I), a human serum-derived protein, recently demonstrated to activate key pluripotency pathways in mouse PS cells. I?I efficiently induces attachment and long-term growth of both embryonic and induced hPS cell lines when added as a soluble protein to the medium at seeding. I?I supplementation efficiently supports adaptation of feeder-dependent hPS cells to xeno-free conditions, clonal growth as well as single-cell survival in the absence of Rho-associated kinase inhibitor (ROCKi). This time-efficient and simplified culture method paves the way for large-scale, high-throughput hPS cell culture, and will be valuable for both basic research and commercial applications.

Citation

Pijuan-Galitó, S., Tamm, C., Schuster, J., Sobol, M., Forsberg, L., Merry, C. L., & Annerén, C. (2016). Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture. Nature Communications, 7, 1-12. https://doi.org/10.1038/ncomms12170

• Journal Article Type: Article
• Acceptance Date: Jun 3, 2016
• Online Publication Date: Jul 13, 2016
• Publication Date: Jul 13, 2016
• Deposit Date: Apr 25, 2017
• Publicly Available Date: Mar 26, 2020
• Journal: Nature Communications
• Electronic ISSN: 2041-1723
• Publisher: Nature Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 7
• Article Number: 12170
• Pages: 1-12
• DOI: https://doi.org/10.1038/ncomms12170
• Public URL: https://nottingham-repository.worktribe.com/output/1114871
• Publisher URL: https://www.nature.com/articles/ncomms12170
• PMID: 27405751