Kawase M, Miura N, Kurikawa N, Masuda K, Higashiyama S, Yagi K, Mizoguchi T
Faculty of Agriculture, Kagawa University, Takamatsu, Japan.
Biol Pharm Bull. 1999 Sep;22(9):999-1001. doi: 10.1248/bpb.22.999.
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 increased the number of attached hepatocytes.
一种三肽生长因子，甘氨酰-L-组氨酰-L-赖氨酸（GHK），被固定在聚乙烯醇季铵化芪唑（PVA-SbQ）凝胶表面。使用光反应性物质4-(3-三氟甲基氮杂环丙基)苯甲酰基-N-羟基琥珀酰亚胺（TDBA-OSu）来连接凝胶和配体GHK。固定化GHK的密度为70 nmol/cm²。将分离的大鼠肝细胞接种到固定有GHK的PVA-SbQ凝胶上并培养5天。接种后约24小时，肝细胞开始聚集并形成多细胞球体，而几乎没有细胞附着在未固定GHK的PVA-SbQ凝胶上。形成的球体在PVA-SbQ凝胶表面牢固附着5天。因此，GHK被证明是肝细胞附着的有效配体。使用十二亚甲基二胺来延长凝胶表面与GHK之间的长度。长度的延长显著增加了附着的肝细胞数量。
