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大鼠胰星状细胞体外分离培养的实验研究

Experimental Study on Isolation and Culture of Pancreatic Stellate Cells in Rats

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【作者】 万远太; 赵秋; 王天才; 王波; 李德明;

【Author】 Wan Yuantai, Zhao Qiu, Wang Tiancai et al Institute of Liver Diseases,Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030

【机构】 华中科技大学同济医学院附属同济医院肝病研究所; 华中科技大学同济医学院附属同济医院肝病研究所 武汉430030; 武汉430030; 武汉430030;

【摘要】 目的 体外分离培养胰星状细胞 (PSC) 以建立合适的体外模型, 为进一步研究胰纤维化的分子发生机制打下实验基础。方法 采用改良Apte MV酶消化法体外分离培养PSC, 胶质原纤维酸性蛋白 (GFAP) 免疫组化染色和荧光倒置显微镜观察自发绿色荧光对原代培养的 PSC加以鉴定。结果 PSC与肝星状细胞 (HSC) 具有相似的特征, 细胞呈梭形或星形, 细胞骨架蛋白 GFAP染色阳性, 由于胞质内含维生素 A脂滴, 在荧光倒置显微镜下于 328nm波长处可见自发绿色荧光。结论 改良酶消化法可用于体外分离培养 PSC, PSC分离培养的成功为进一步研究PSC活化与抑制信号通路的分子机制提供了体外模型。

【Abstract】 Objective To establish a method for the isolation and culture of pancreatic stellate cells (PSCs) in rats. Methods PSCs were isolated and cultured by using modified enzyme digestive method with Apte MV, and examined by immunostaining for glial fibrillary acidic protein (GFAP). The PSCs of primary culture were identified by observing the spontaneous green fluorescence under the fluorescence inverted microscopy. Results The rat PSCs exhibited features similar to those described for hepatic stellate cells (HSCs). PSCs had a flattened angular appearance or stellate shape. The immunostaining of the GFAP was positive. Spontaneous green fluorescence was observed under the fluorescence inverted microscopy at 328 nm due to cytomplasma-containing vitamin A lipid droplets. Conclusion PSCs were isolated and cultured successfully by modified enzyme digestive method.

  • 【文献出处】 华中科技大学学报(医学版) ,Acta Universitatis Medictnae Tangji , 编辑部邮箱 ,2005年01期
  • 【分类号】R329.2
  • 【被引频次】5
  • 【下载频次】103
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