RRC ID 42781
著者 Watanabe K, Yaguchi T, Yang D, Kanno T, Nagai K, Yamamoto S, Fujikawa H, Yamamoto H, Nagata T, Tashiro C, Nishizaki T.
タイトル Beneficial effect of intracellular free high-mannose oligosaccharides on cryopreservation of mammalian cells and proteins.
ジャーナル Cryobiology
Abstract The cryoprotective effect of intracellular free high-mannose oligosaccharides (HMOS) on mammalian cells and proteins was examined by monitoring PC-12 cell viability and assaying protein kinase C (PKC)-epsilon activity. 1-Deoxymannojirimycin, an inhibitor of alpha-mannosidase, to cause an increase in intracellular free HMOS, significantly rescued PC-12 cells with 2-h freezing insult at -15 degrees C in a concentration (1-50mM)- and pretreatment time (48-72h)-dependent manner, as compared with unpretreated cells; full rescue from freezing injury was obtained with 1-deoxymannojirimycin at more than 25mM for 48-h pretreatment and more than 3mM for 72- and 96-h pretreatment. For PC-12 cells pretreated with 1-deoxymannojirimycin at 1mM for 72h, thawed cell viability after more than 8-w cryopreservation at -80 degrees C in 10% (v/v) dimethyl sulfoxide was much higher than that for cells without pretreatment. PKC-epsilon activity was well preserved after 16-h cryopreservation at -20 degrees C in the presence of mannose 9-N-acetylglucosamine 2 (Man9-GlcNAc2) (1 mM), an HMOS, while the activity was reduced to 15% without Man9-GlcNAc2. Collectively, the results of the present study suggest that intracellular free HMOS is a key molecule to protect mammalian cells and proteins from freezing injury; in other words, HMOS could be a new target for cryopreservation of mammalian cells and proteins.
巻・号 53(3)
ページ 330-5
公開日 2006-12-1
DOI 10.1016/j.cryobiol.2006.08.004
PII S0011-2240(06)00119-2
PMID 17026979
MeSH 1-Deoxynojirimycin / pharmacology Animals Cell Survival / drug effects Cryopreservation* Cryoprotective Agents / pharmacology Enzyme Inhibitors / pharmacology Intracellular Fluid / metabolism Mannans / metabolism* Mannans / pharmacology Mannose / chemistry Oligosaccharides / chemistry Oligosaccharides / metabolism* PC12 Cells Protein Kinase C-epsilon / metabolism Rats alpha-Mannosidase / antagonists & inhibitors
IF 2.287
引用数 2
WOS 分野 BIOLOGY PHYSIOLOGY
リソース情報
ヒト・動物細胞