title: Differentiation of oligodendrocytes from mouse induced pluripotent stem cells without serum.
abstract: Cell therapy using induced pluripotent stem (iPS) cells might become a new approach for treating neonatal hypoxic-ischemic injury such as periventricular leukomalacia. To obtain appropriate donor cells for transplantation, we differentiated oligodendrocyte (OL) lineage cells from mouse iPS cells. Induction of OL lineage cell differentiation from iPS cells was carried out with a seven-step culture method. Mouse iPS cells (stage 1) were induced to form embryoid bodies for 4 days under a serum-free condition that was suitable for ectoderm induction (stage 2), following by selection of nestin-positive neural stem cells (NSCs) for 10-12 days (stage 3). NSCs were cultured in expansion medium containing fibroblast growth factor (FGF)-2 for 4 days (stage 4), induced to differentiate into glial progenitor cells by epidermal growth factor and fibroblast growth factor (FGF-2) treatment for 4-5 days (stage 5), and then into OL progenitor cells by culture in neurobasal A medium containing FGF-2 and platelet-derived growth factor for 6-8 days (stage 6). Terminal differentiation into O4-positive OLs was carried out by culture in neurobasal A containing T3 and ciliary neurotrophic factor for 7 days (stage 7). Inwardly rectifying K+ currents, which are characteristic of OLs, were detected in iPS cell-derived cells at stage 7 in whole cell clamp mode. Our data suggest that OLs can be effectively differentiated from mouse iPS cells without serum in a stepwise manner, which may be appropriate for use as donor cells in transplantation.
authors: Department of Neurophysiology and Brain Science, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, 467-8601, Japan.
authors: Sachiyo
authors: S
authors: Misumi
authors: Ruriko
authors: R
authors: Nishigaki
authors: Yoshitomo
authors: Y
authors: Ueda
authors: Yoko
authors: Y
authors: Watanabe
authors: Yuko
authors: Y
authors: Shimizu
authors: Akimasa
authors: A
authors: Ishida
authors: ChaGyun
authors: C
authors: Jung
authors: Hideki
authors: H
authors: Hida
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D002469:Cell Separation; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice; D009836:Oligodendroglia / Q000166:cytology* / Q000502:physiology; D018408:Patch-Clamp Techniques; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction