Document Detail

Beneficial effect of intracellular free high-mannose oligosaccharides on cryopreservation of mammalian cells and proteins.
MedLine Citation:
PMID:  17026979     Owner:  NLM     Status:  MEDLINE    
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.
Kanako Watanabe; Takahiro Yaguchi; Dongqin Yang; Takeshi Kanno; Kaoru Nagai; Satoshi Yamamoto; Hirokazu Fujikawa; Hideyuki Yamamoto; Tetsu Nagata; Chikara Tashiro; Tomoyuki Nishizaki
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Publication Detail:
Type:  Journal Article     Date:  2006-10-05
Journal Detail:
Title:  Cryobiology     Volume:  53     ISSN:  0011-2240     ISO Abbreviation:  Cryobiology     Publication Date:  2006 Dec 
Date Detail:
Created Date:  2007-01-08     Completed Date:  2007-03-07     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  330-5     Citation Subset:  IM    
Department of Physiology, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan.
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MeSH Terms
1-Deoxynojirimycin / pharmacology
Cell Survival / drug effects
Cryoprotective Agents / pharmacology
Enzyme Inhibitors / pharmacology
Intracellular Fluid / metabolism
Mannans / metabolism*,  pharmacology
Mannose / chemistry
Oligosaccharides / chemistry,  metabolism*
PC12 Cells
Protein Kinase C-epsilon / metabolism
alpha-Mannosidase / antagonists & inhibitors
Reg. No./Substance:
0/Cryoprotective Agents; 0/Enzyme Inhibitors; 0/Mannans; 0/Oligosaccharides; 0/mannosyl(9)-N-acetylglucosamine2; 19130-96-2/1-Deoxynojirimycin; 31103-86-3/Mannose; EC Kinase C-epsilon; EC

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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