大鼠肝卵圆细胞的分离培养和体外分化研究  

Isolation,culture and in vitro differentiation of rat oval cells

作　　者：单云峰[1] 周伟平[1] 杨文[1] 刘淑琴[1] 曹惠芳[1] 邱秀华[1] 吴孟超[1] 王红阳[1] 

机构地区：[1]第二军医大学东方肝胆外科医院外科

出　　处：《中华实验外科杂志》2005年第8期925-926,i0003,共3页Chinese Journal of Experimental Surgery

基　　金："973"国家重点基础研究发展规划项目(2004CB518703)

摘　　要：目的观察大鼠肝卵圆细胞的活化,为进一步研究肝卵圆细胞特征建立简单的分离培养方法。方法利用2乙酰氨基芴/部分肝切除建立肝卵圆细胞活化的大鼠模型,采用选择性消化法从该模型中分离纯化肝卵圆细胞,并做免疫细胞荧光鉴定。用丁酸钠诱导其分化。结果成功地建立了肝卵圆细胞活化模型,并从中分离培养了表达OV6、CK19、AFP、白蛋白、ckit、Thy1、CD45和CD34的卵圆细胞。在丁酸钠刺激下它可向肝细胞分化。结论利用选择性消化法可以分离到肝卵圆细胞,并且其具有肝干细胞的特征。Objective To observe the activation of oval cells in adult rat liver and to enrich them for further analysis of characterization in vitro, Methods Rat model for hepatic oval cell proliferation was established with 2-acetylaminofluorene and two third partial hepatectomy （2-AAF/PH）. Paraffin-embedded rat liver sections from model （7 days after hepatectomy） and control groups were stained with HE and OV6 antibodies by immunohistochemistry. The oval cells were isolated in this model, cultured for different digest time and cofirmed by immunofluorescence assay. Differentiatiort was induced by sodium butyrate. Results A 2-AAF/PH model was successfully established to activate the oval cell compartment in rat liver. The sorted oval cells formed colonies when cultured in vitro. Cells in the colony expressed OV6,CK19, AFP, albumin, c-kit, Thyl, CD45 and CD34. After sodium butyrate was added in the medium, the oval cells differentiated into liver cells. Conclusion Hepatic oval cells can be isolated from the 2-AAF/PH model by different digest time and they had the properties of hepatic stem/progenitor cells.

关 键 词：大鼠 肝卵圆细胞 细胞分离 细胞培养 体外分化 丁酸钠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]