大鼠肝星状细胞的培养改良与性质鉴定

发布时间：2018-06-17 15:28

本文选题：大鼠肝星状细胞 + 培养　；参考：《临床医药文献电子杂志》2014年10期

【摘要】：目的肝星状细胞(hepatic stellate cell,HSC)是一类胞浆内富含Vitamin A脂滴的特殊类型的细胞,激活后的HSC胞浆中Vitamin A脂滴减少活消失,是研究肝纤维化疾病的良好素材。本研究欲改良HSC细胞的培养方法,并鉴别四种HSC细胞鉴定方法的有效性。方法采用含0.2 m M油酸的完全培养基对已培养传代的大鼠肝星状细胞进行培养。分别采用油红0染色、Vitamin A脂滴的紫外激发、a-SMA免疫组化、a-SMA免疫荧光四种方法对培养的HSC进行鉴定。结果用含0.2 m M油酸的完全培养基培养的大鼠肝星状细胞,细胞形态饱满,胞浆不因传代而减少,细胞形态维持稳定,油红O染色后可清晰显示未激活HSC内的脂滴;紫外激发下Vitamin A脂滴发出极易淬灭的蓝绿色荧光;以泡球蚴囊液为外源性刺激,使HSC激活,a-SMA免疫组化染色激活的HSC胞浆均可见棕黄色颗粒状沉淀;a-SMA免疫荧光显示激活的HSC细胞胞浆均可见FITC的激发的绿色荧光。结论含0.2 m M油酸的完全培养基可有效维持细胞形态。油红0染液、Vitamin A脂滴的紫外激发可有效鉴定未激活的HSC,a-SMA免疫组化、a-SMA免疫荧光方法是鉴别激活的HSC的有效方法。
[Abstract]:Objective Hepatic stellate cell (HSC) is a special type of cells rich in Vitamin A lipid droplets in the cytoplasm. The reduced activity of Vitamin A lipid droplets in activated HSCs is a good material for the study of hepatic fibrosis. The aim of this study was to improve the culture method of HSC cells and to identify the effectiveness of four HSC cell identification methods. Methods cultured rat hepatic stellate cells were cultured on a complete culture medium containing 0.2 mm oleic acid. The cultured HSC were identified by oil red 0 staining and ultraviolet stimulated SMA immunofluorescence staining. Results Rat hepatic stellate cells cultured in 0.2 mm oleic acid culture medium were full in shape, no decrease in cytoplasm due to passage, cell morphology remained stable, lipid droplets in HSC could be clearly displayed after oil red O staining. Vitamin A lipid droplets emitted easily quenched blue-green fluorescence under ultraviolet excitation. The cytoplasm of HSC activated by HSC was stained by immunohistochemical staining. All the HSC cells activated by HSC showed the green fluorescence induced by FITC in the cytoplasm of HSC activated by brown granular precipitate and a-SMA immunofluorescence. Conclusion the complete culture medium containing 0.2 mm oleic acid can effectively maintain cell morphology. Ultraviolet excitation of Vitamin A lipids in oil red 0 dye solution can effectively identify the unactivated HSCA- SMA immunofluorescence method is an effective method to identify activated HSC.
【作者单位】：青海大学附属医院肝胆胰外科;
【分类号】：R329.2

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