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

• Published in: Nature communications Vol. 7; no. 1; p. 12170
• Main Authors: Pijuan-Galitó, Sara, Tamm, Christoffer, Schuster, Jens, Sobol, Maria, Forsberg, Lars, Merry, Catherine L. R., Annerén, Cecilia
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 13.07.2016; Nature Publishing Group; Nature Portfolio
• Subjects: 101/58; 13; 13/1; 13/100; 13/106; 13/51; 14; 14/19; 14/63; 38; 38/22; 38/39; 38/77; 38/90; 631/136/532/2064; 631/1647/1407/651; 631/45/612/1221; 631/80; 82; 82/29; Alpha-Globulins; Cell culture; Cell Culture Techniques - methods; Cell Proliferation; Cells, Cultured; Culture Media; Human Embryonic Stem Cells - cytology; Humanities and Social Sciences; Humans; Induced Pluripotent Stem Cells - cytology; Kinases; multidisciplinary; Pluripotent Stem Cells - cytology; Proteins; Science; Science (multidisciplinary); Stem cells; Sweden
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms12170

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-α-inhibitor (IαI), a human serum-derived protein, recently demonstrated to activate key pluripotency pathways in mouse PS cells. IαI 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. IαI 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. Improved culture methods are needed to reliably grow human pluripotent stem cells (hPSCs) on a large scale. Here, the authors identify a xeno-free medium with a supplement of Inter-α-inhibitor that supports long-term propagation and improved single-cell passaging of hPSCs on uncoated plastic.