大鼠肝内淋巴细胞的分离与培养

Isolation and culture of rat intrahepatic lymphocytes

目的建立一种简单易行的大鼠肝内原代淋巴细胞分离与培养方法,为肝脏免疫学研究提供可靠的体外细胞模型。方法采用原位灌注及密度梯度离心法分离培养,即在无菌条件下取大鼠正常肝脏组织,研磨后,利用离心法及淋巴细胞分离液进行纯化,通过倒置生物显微镜对所培养细胞进行形态学观察,并结合流式细胞术来鉴定大鼠T淋巴细胞。结果通过倒置生物显微镜观察到细胞呈典型的淋巴细胞形态,流式细胞术分析显示,T细胞比例为51.2%。结论本实验利用原位灌注及密度梯度离心法相结合的方式,成功分离和培养了大鼠肝内淋巴细胞,操作简单,重复性好,成功率高,为进一步开展肝脏的免疫学研究提供了非常好的体外细胞模型。

Objective To establish a simple and convenient method for isolation and culture of primary intrahepatic lymphocytes,and to provide a reliable cell model for liver immunological study.Methods Situ perfusion and density gradient centrifugation were used for isolation and culture,the liver tissue of rat was removed under aseptic condition and grinded and purified by centrifugation and lymphocyte isolation solution;morphological changes of cultured cells were observed by inverted biological microscope,and T lymphocyte was identified by flow cytometry.Results The typical lymphocyte morphology was observed by inverted biological microscope,and the proportion of T cells was 51.2%by flow cytometry.Conclusion In this study,intrahepatic lymphocytes were successfully isolated and cultured by situ perfusion and density gradient centrifugation,the method is uncomplicated,reproducible and has high success rate.It provides a beyond compare cell model for further study of liver immunology.