摘要
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。
AIM To observe the phenotypic activation and c-fos, c-jun expression of in vitro cultured human hepatic stellate cells (HSCs).METHODS HSCs isolated from human livers were cultured and subcultured in DMEM containing calf serum. Primary and secondary cell cultures were viewed under phase-contrast microscopy and PCNA, procollagen Ⅰ , α-SMA, c-fos and c-jun protein expression of HSCs in primary and secondary cultures were detected by immunocytochemical staining.RESULTS Human HSCs cultured in DMEM containing 10% calf serum under went phenotypic changes and transdifferentiated from a resting cell during early days of primary culture into a activated cell during late days of primary culture or after subculture. Activated human HSCs were associated with characteristic myofibroblast-like morphology and obvious immunocytochemical staining for PCNA, procollagen Ⅰ, and α-SMA. c-fos and c-jun expression of primary HSCs cultured in DMEM without calf serum for 24 hours were negative and became positive when cultured for another 24 hours in medium containing 10% calf serum. Positive c-fos and c-jun expression was also observed on HSCs in primary culture for 10 days and at the third day of first passage. CONCLUSION Human HSCs cultured in serum containing medium spontaneously undergo phenotypic activation and increased nuclear expression of c-fos and c-jun may be involved in the phenotypic activation.
出处
《世界华人消化杂志》
CAS
2000年第3期299-302,共4页
World Chinese Journal of Digestology
关键词
肝
细胞学
星形细胞
蛋白表达
免疫细胞化学
liver/cytology
stellate cell
cells, cultured
c-fos
c-jun
protein expression
immunocytochemistry
