RRC ID 44056
Author Tomizawa M, Shinozaki F, Sugiyama T, Yamamoto S, Sueishi M, Yoshida T.
Title Activin A is essential for Feeder-free culture of human induced pluripotent stem cells.
Journal J Cell Biochem
Abstract Feeder-free culture of human induced pluripotent stem (hiPS) cells is necessary for their clinical application to avoid adverse effects of foreign proteins. hiPS cells were cultured with combinations of activin (A), CHIR99021 (C), basic fibroblast growth factor (F), and leukemia inhibitory factor (L) under feeder-free conditions. Culture was terminated after 12 passages or when the cell morphology changed from pluripotency. Pluripotency was analyzed by alkaline phosphatase (ALP) staining and immunostaining with antibodies to Oct3/4, Nanog, SSEA4, and TRA-1-60. SB431542 (SB), an activin inhibitor, was added to the culture, and the morphology of the cells was observed. hiPS cells cultured with A, AC, and ACL after 12 passages were positive for ALP staining. Oct3/4 was positive in hiPS cells cultured with A, AC, and ACL. hiPS cells were positive for Nanog when cultured with A and AC; however, Nanog signal was weaker in cells cultured with ACL. SSEA4 was positive in hiPS cells cultured with A and AC but almost negative in those cultured with ACL. hiPS cells were positive for TRA-1-60 when cultured with A, AC, and ACL. hiPS cells lose their undifferentiated morphology at six passages when cultured with A + SB, five passages with AC + SB, and nine passages with ACL. We conclude that feeder-free culture of hiPS cells requires A or AC to maintain pluripotency.
Volume 114(3)
Pages 584-8
Published 2013-3
DOI 10.1002/jcb.24395
PMID 22991093
MeSH Activins / antagonists & inhibitors Activins / pharmacology* Alkaline Phosphatase / analysis Animals Antigens, Surface / analysis Benzamides / pharmacology Cell Culture Techniques* Collagen Dioxoles / pharmacology Drug Combinations Feeder Cells* Fibroblast Growth Factor 2 / pharmacology Homeodomain Proteins / analysis Humans Induced Pluripotent Stem Cells / metabolism* Laminin Leukemia Inhibitory Factor / pharmacology Mice Nanog Homeobox Protein Octamer Transcription Factor-3 / analysis Proteoglycans / analysis Pyridines / pharmacology* Pyrimidines / pharmacology* Stage-Specific Embryonic Antigens / analysis
IF 3.448
Times Cited 6
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY
Resource
Human and Animal Cells