论文部分内容阅读
目的 :观察常规培养条件下正常人肝脏星形细胞 (HSCs)表型激活并对其进行鉴定。方法 :由正常人肝脏中分离HSCs ,在未包被的玻璃及塑料培养支持物上 ,以常规的含小牛血清的DMEM培养 ,相差显微镜下观察培养过程中细胞形态改变 ,荧光显微镜下观察维生素A自发荧光 ,免疫细胞化学染色观察α SMA、Ⅰ型胶原、FN及PCNA改变。结果 :正常人HSCs未包被的玻璃及塑料培养支持物上培养时 ,其表型由刚分离时及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样形态 ,胞质中无明显脂滴 ,维生素A自发荧光明显减弱或消失 ,同时明显表达α SMA、Ⅰ型胶原、FN及PCNA。结论 :体外常规培养可诱导人HSCs激活 ,激活的人HSCs具有典型的形态及功能特点。
Objective: To observe the phenotype activation of normal human hepatic stellate cells (HSCs) under routine culture conditions and to identify them. Methods: HSCs were isolated from normal human liver, cultured on uncoated glass and plastic culture support, cultured in conventional DMEM containing calf serum, observed morphological changes during culture under phase contrast microscope, observed the vitamins under the fluorescence microscope A autofluorescence, immunocytochemical staining α SMA, type Ⅰ collagen, FN and PCNA changes. Results: When HSCs were cultured on glass and plastic substrates, the phenotypes of normal HSCs were changed from resting type at the beginning of primary culture to the later stage of primary culture and the activated type after passage. Activated human HSCs showed a typical fibroblast-like morphology with no lipid droplets in the cytoplasm. Autofluorescence of vitamin A significantly decreased or disappeared, while α SMA, type I collagen, FN and PCNA were also expressed. CONCLUSION: Human HSCs can be induced to activate in normal culture in vitro. The activated human HSCs have typical morphological and functional characteristics.