RRC ID |
42781
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著者 |
Watanabe K, Yaguchi T, Yang D, Kanno T, Nagai K, Yamamoto S, Fujikawa H, Yamamoto H, Nagata T, Tashiro C, Nishizaki T.
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タイトル |
Beneficial effect of intracellular free high-mannose oligosaccharides on cryopreservation of mammalian cells and proteins.
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ジャーナル |
Cryobiology
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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.
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巻・号 |
53(3)
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ページ |
330-5
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公開日 |
2006-12-1
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DOI |
10.1016/j.cryobiol.2006.08.004
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PII |
S0011-2240(06)00119-2
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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
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引用数 |
2
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WOS 分野
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BIOLOGY
PHYSIOLOGY
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リソース情報 |
ヒト・動物細胞 |
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