成体小鼠肝脏卵圆细胞的分离及培养  

Isolation and culture of hepatic oval cells from adult mice

作　　者：张磊[1] 万伟伟[1] 梁慧芳[1] 陈孝平[1] 张万广[1] 张伟[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝脏外科中心,湖北武汉430030

出　　处：《中国普通外科杂志》2008年第11期1084-1087,共4页China Journal of General Surgery

基　　金：国家自然科学基金重点资助项目(30430670);卫生部部属医院临床学科重点项目(2007-353);湖北省肝脏外科医学研究中心项目

摘　　要：目的建立一种稳定的成体小鼠卵圆细胞的分离和培养方法。方法采用喂饲2-乙酰氨基芴(AAF)加2/3肝切除方法诱导肝脏卵圆细胞的增殖。经门静脉灌注消化法和等密度离心法分离卵圆细胞,并在体外进行长期培养。免疫荧光和免疫双标法对培养的卵圆细胞加以鉴定。结果体外培养的卵圆细胞呈集落样生长,稳定传代并已培养至3个月。免疫荧光和免疫双标法证实培养的细胞为卵圆细胞,并显示该细胞具有分化潜能。结论该方法是一种稳定的成体小鼠卵圆细胞的分离和培养方法,为肝脏干细胞的相关研究和应用奠定基础。Objective To establish a reliable method for isolation and culture of murine adult oval cells. Methods Oval cell response was stimulated by treatment with N-2-aeetylaminofluorene （2-AAF） diet and partial hepatectomy. A modification of a two-step perfusion protocol and isopycnie centrifugation were used to isolate oval cells from livers. Immunoftuoreseenee and double-fluorescence immunostaining were used for identification of oval cells. Results Isolated oval cells were grown as colonies in vitro and had been cuhured for three months. Isolated cells were identification by immunofluoreseence and double-fluorescence immunostaining, which also indicated isolated cells were bipotential. Conclusions This method mentioned above is a reliable method for isolation and primary culture of oval cells, which would provide a useful tool for investigating oval cell biology.

关 键 词：卵圆细胞 肝 细胞分离 细胞鉴定 细胞培养 

分 类 号：R36-33[医药卫生—病理学]