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目的 观察体外培养过程中人肝脏星形细胞(HSCs)表型及其c-fos和c-jun表达的改变。 方法 将分离的正常人HSCs进行原代及传代培养,倒置显微镜下动态观察培养细胞形态改变,对原代及传代培养细胞铺展片进行PCNA、Ⅰ型前胶原、α-SMA,c-fos及c-jun免疫细胞化学染色。 结果 正常人HSCs在含100mL/L小牛血清中培养时,其表型由原代培养初期的静息型转变为原代培养后期及传代后的激活型,激活的人HSCs呈现典型的成纤维细胞形态特征,其表达PCNA、工型前胶原及a-SMA明显阳性,刚分离的正常人HSCs在不含血清的培养液中培养24h后,其c-fos及c-jun表达均为阴性,而在含100mL/L小牛血清的培养液中继续培养24h后,c-fos及c-jun表达为阳性,原代培养d10及传代培养d3的HSCs其c-fos及c-jun表达持续阳性。 结论 在含小牛血清的DMEM培养液中培养时,人HSCs自发地激活,这种激活可能与c-fos及c-jun表达增加有关。
Objective To observe the phenotype of human hepatic stellate cells (HSCs) and the expression of c-fos and c-jun in vitro culture. Methods The HSCs isolated from normal individuals were subcultured in primary culture and subculture. The morphological changes of cultured HSCs were observed under inverted microscope. PCNA, type Ⅰ procollagen, α-SMA, c-fos and c -jun immunocytochemical staining. Results When HSCs were cultured in 100 mL / L fetal bovine serum, the phenotype of normal HSCs changed from resting type to primary culture type and post-passage type. The activated HSCs presented typical fibroblasts The morphological features of PCNA, PCNA and type-A procollagen were significantly positive. The normal HSCs isolated from normal HSCs cultured in serum-free medium for 24 hours showed negative expression of c-fos and c-jun, However, the expression of c-fos and c-jun was positive in the culture medium containing 100mL / L fetal bovine serum for 24 hours, while the expression of c-fos and c-jun in the primary cultured d10 and subcultured d3 HSCs remained positive . Conclusions Human HSCs are activated spontaneously in DMEM medium containing fetal bovine serum, which may be related to the increased expression of c-fos and c-jun.