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

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.

SUBMITTER: Pijuan-Galito S 

PROVIDER: S-EPMC4947164 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature