Back to Search Start Over

Beneficial effect of intracellular free high-mannose oligosaccharides on cryopreservation of mammalian cells and proteins.

Authors :
Watanabe K
Yaguchi T
Yang D
Kanno T
Nagai K
Yamamoto S
Fujikawa H
Yamamoto H
Nagata T
Tashiro C
Nishizaki T
Source :
Cryobiology [Cryobiology] 2006 Dec; Vol. 53 (3), pp. 330-5. Date of Electronic Publication: 2006 Oct 05.
Publication Year :
2006

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.

Details

Language :
English
ISSN :
0011-2240
Volume :
53
Issue :
3
Database :
MEDLINE
Journal :
Cryobiology
Publication Type :
Academic Journal
Accession number :
17026979
Full Text :
https://doi.org/10.1016/j.cryobiol.2006.08.004