title: A neurosphere-derived factor, cystatin C, supports differentiation of ES cells into neural stem cells.
abstract: Although embryonic stem (ES) cells are capable of unlimited proliferation and pluripotent differentiation, effective preparation of neural stem cells from ES cells are not achieved. Here, we have directly generated under the coculture with dissociated primary neurosphere cells in serum-free medium and the same effect was observed when ES cells were cultured with conditioned medium of primary neurosphere culture (CMPNC). ES-neural stem cells (NSCs) could proliferate for more than seven times and differentiate into neurons, astrocytes, and oligodendrocytes in vitro and in vivo. The responsible molecule in CMPNC was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, which turned out to be cystatin C. Purified cystatin C in place of the CMPNC could generate ES-NSCs efficiently with self-renewal and multidifferentiation potentials. These results reveal the validity of cystatin C for generating NSCs from ES cells.
authors.affiliation: Department of Pediatrics, Horizontal Medical Research Organization, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Takeo
authors.initials: T
authors.lastname: Kato
authors.forename: Toshio
authors.initials: T
authors.lastname: Heike
authors.forename: Katsuya
authors.initials: K
authors.lastname: Okawa
authors.forename: Munetada
authors.initials: M
authors.lastname: Haruyama
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Shiraishi
authors.forename: Momoko
authors.initials: M
authors.lastname: Yoshimoto
authors.forename: Masako
authors.initials: M
authors.lastname: Nagato
authors.forename: Minoru
authors.initials: M
authors.lastname: Shibata
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kumada
authors.forename: Yasunari
authors.initials: Y
authors.lastname: Yamanaka
authors.forename: Haruo
authors.initials: H
authors.lastname: Hattori
authors.forename: Tatsutoshi
authors.initials: T
authors.lastname: Nakahata
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D018920:Coculture Techniques; D003342:Corpus Striatum / Q000196:embryology; D017077:Culture Media, Conditioned; D055316:Cystatin C; D015891:Cystatins / Q000502:physiology*; D004622:Embryo, Mammalian; D005434:Flow Cytometry; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D013234:Stem Cells / Q000166:cytology*
chemical_list: C525031:Cst3 protein, mouse; D017077:Culture Media, Conditioned; D055316:Cystatin C; D015891:Cystatins