纤维连接蛋白调理的人表皮角质细胞粘附、迁移功能

目的观察纤维连接蛋白(FN)对人表皮角质细胞(EKC)在体外培养过程中有无粘附、迁移作用.方法利用不同剂量的FN,对新鲜EKC及体外培养7～10d的EKC进行粘附、迁移功能测定.应用间接免疫荧光染色法,检测α5β1受体在细胞培养前后的表达.结果(1)新鲜EKC在FN调理下的粘附、迁移值明显低于体外培养7～10d的EKC,粘附值分别为(0.9±0.5)%、(37.26±8.34)%(P＜0.01),迁移指数分别为17.5±2.5、48.7±3.2(P＜0.01).(2)培养1d的EKC及组织块体外培养后增生的表皮细胞,α5β1受体的表达染色阳性;培养7d的EKC及组织块增生表皮外周边缘染色呈强阳性;新鲜EKC染色阴性或弱阳性.结论(1)新鲜EKC与培养7～10d的EKC生物学功能间差异有显著性意义.(2)EKC生物学功能活跃的阶段(培养7d)和部位(组织块边缘增生的表皮细胞)α5β1受体表达最强.(3)体外培养能有效地诱导和激活EKC的粘附、迁移功能,使生物学功能相对静止的EKC变成了功能活跃的EKC细胞.

The adhesive and migrating function of human epithelial cells opsonized by fibronectin

Objective To investigate the adhesive and migrating function of human epithelial keratio- cytes during in vitro culture. Methods The adhesive and migrating function was determined in freshly iso- lated EKCs and those cultured for 7 ~ 10 days after being opsonized by fibronectin( FN). The expressions of a5P, receptors in EKC were examined with indirect immunofluorescence staining methods before and after culture. Results ( 1) The adhesive and migrating indices after FN opsonization of EKCs freshly isolated EKCs were obviously lower than those cultured for 7 ~ 10 days ( P < 0. 01). (2) There exhibited positive staining of the expresson of a5P, receptors in the EKC after 1 day culture and in the proliferative epithelia af- ter in vitro culturing of a tissue mass. While there exhibited strongly positive staining of the peripheral prolif- erative epithelia of the EKCs and tissue mass after 7 days of culture, negative or weakly positive stainings were found in freshly isolated EKCs. Conclusion ( 1 ) There existed significant difference of biological function between the freshly isolated EKCs and those cultured for 7 ~ 10 days. (2) The strongest expression of a5p[ receptors was observed at the active proliferative site(peripheral proliferative epithelia of a tissue mass) and in the biological active period (cultured for 7 days) of EKCs. (3) The adhesive and migrating function of EKCs could be effectively induced and activated by in vitro culture, which enabled the EKC to alter from a relatively biologically functional static state to an actively functional state.