A neurosphere-derived factor, cystatin C, supports differentiation of ES cells into neural stem cells.
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.
Kato
Takeo
T
Department of Pediatrics, Horizontal Medical Research Organization, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Heike
Toshio
T
Okawa
Katsuya
K
Haruyama
Munetada
M
Shiraishi
Kazuhiro
K
Yoshimoto
Momoko
M
Nagato
Masako
M
Shibata
Minoru
M
Kumada
Tomohiro
T
Yamanaka
Yasunari
Y
Hattori
Haruo
H
Nakahata
Tatsutoshi
T
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*
C525031:Cst3 protein, mouse; D017077:Culture Media, Conditioned; D055316:Cystatin C; D015891:Cystatins