Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
Immobilization of Tripeptide Growth Factor Glycyl-L-histidyl-L-lysine on Poly(vinylalcohol)-quarternized Stilbazole (PVA-SbQ) and Its Use as a Ligand for Hepatocyte Attachment
Masaya KAWASENagamitsu MIURANobuya KURIKAWAKaori MASUDAShinji HIGASHIYAMAKiyohito YAGITadashi MIZOGUCHI
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1999 Volume 22 Issue 9 Pages 999-1001

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Abstract

A tripeptide growth factor, glycyl-L-histidyl-L-lysine (GHK), was immobilized on the surface of poly(vinylalcohol)-quarternized stilbazole (PVA-SbQ) gel. The photoreactive substance, 4-(3-trifluoromethylazirino)benzoyl-N-hydroxysuccinimide (TDBA-OSu), was employed to link the gel and ligand GHK. The density of immobilized GHK was 70 nmol/cm2. Isolated rat hepatocytes were inoculated on the GHK-immobilized PVA-SbQ gel and cultured for 5 d. About 24 h after inoculation, hepatocytes started to aggregate and formed multicellular spheroids while almost no cells attached to GHK-non-immobilized PVA-SbQ gel. The formed spheroids attached firmly to the surface of PVA-SbQ gel for 5 d. GHK was, thus, shown to be an effective ligand for hepatocyte attachment. Dodecamethylenediamine was used to extend the length between the gel surface and GHK. Extension of the length significantly increaesd the number of attached hepatocytes.

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