title: 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-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.
authors.affiliation: Department of Medical Biochemistry and Microbiology, Box 582, Uppsala University, 751 23 Uppsala, Sweden.
authors.forename: Sara
authors.initials: S
authors.lastname: Pijuan-Galitó
authors.affiliation: Department of Medical Biochemistry and Microbiology, Box 582, Uppsala University, 751 23 Uppsala, Sweden.
authors.forename: Christoffer
authors.initials: C
authors.lastname: Tamm
authors.affiliation: Department of Immunology, Genetics and Pathology and Science for Life Laboratory, Box 815, Uppsala University, 751 08 Uppsala, Sweden.
authors.forename: Jens
authors.identifier: 0000-0002-4383-9880
authors.initials: J
authors.lastname: Schuster
authors.affiliation: Department of Immunology, Genetics and Pathology and Science for Life Laboratory, Box 815, Uppsala University, 751 08 Uppsala, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Sobol
authors.affiliation: Department of Immunology, Genetics and Pathology and Science for Life Laboratory, Box 815, Uppsala University, 751 08 Uppsala, Sweden.
authors.forename: Lars
authors.initials: L
authors.lastname: Forsberg
authors.affiliation: Department of Medical Biochemistry and Microbiology, Box 582, Uppsala University, 751 23 Uppsala, Sweden.|Stem Cell Glycobiology Group, Wolfson Centre for Stem Cells, Tissue Engineering &Modelling Room A59, University of Nottingham, NG7 2RD Nottingham, UK.
authors.forename: Catherine L R
authors.initials: CL
authors.lastname: Merry
authors.affiliation: Department of Medical Biochemistry and Microbiology, Box 582, Uppsala University, 751 23 Uppsala, Sweden.|GE Healthcare Bio-Sciences AB, Björkgatan 30, 751 84 Uppsala, Sweden.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Annerén
mesh_terms: D000510:Alpha-Globulins; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media; D000066449:Human Embryonic Stem Cells / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology*
chemical_list: D000510:Alpha-Globulins; D003470:Culture Media; C011877:inter-alpha-inhibitor