摘要
本文拟建立大鼠卵圆细胞增殖模型 ,分离纯化卵圆细胞并对其特性进行研究。给SD大鼠喂饲 2 乙酰氨基芴 (2 AAF) ,剂量分别为 5、10、15、2 0mg kg ,连续给药 6d ,第 7天行三分之二肝切除术 ,术后继续给药 1周 ,并于术后每隔 3d取肝组织行常规组织学观察、细胞增殖试验及免疫组化染色。经免疫磁珠标记C kit阳性细胞以纯化卵圆细胞 ,再经免疫细胞化学及RT PCR对其进行鉴定。结果显示 ,10、15、2 0mg kg三种剂量 2 AAF均可成功建立卵圆细胞增殖模型 ,卵圆细胞增殖于第 8天较明显 ,至第 11天达高峰 ,第 14天有所减少。卵圆细胞胞核BrdU染色阳性 ,卵圆细胞表达特异性抗原OV6、肝细胞标记白蛋白、胆管细胞标记CK19、甲胎蛋白以及连接蛋白 4 3,同时还表达造血干细胞标记C kit。实验表明 ,2 AAF剂量为 10～ 2 0mg kg时可获得较理想的大鼠卵圆细胞增殖模型 ,增生的卵圆细胞为具有双向分化潜能的肝干细胞 前体细胞。
This essay was aimed to establish a rat model for hepatic oval cells proliferation and to analyze the characterization of oval cell.SD rats were given 2-acetylaminofluorene (2-AAF)6 days before and 7 days after partial hepatectomy. Liver tissue specimens were obtained every 2～3 days after partial heaptectomy and were investigated by HE staining and immunohistochemical staining for BrdU,OV6, cytokeratin-19,albumin, AFP, connexin-43 and C-kit.Obvious oval cell proliferation was seen in the liver in 10mg/kg,15mg/kg,20mg/kg groups and oval cell BrdU incorporation test was positive. Oval cell expressed its specific marker OV6,hepatic cell marker albumin , biliary cell marker CK19,fetal liver cell marker AFP and connexin-43 as well as hematopoietic stem cell receptor C-kit.It suggests that rat model for hepatic oval cell proliferation can be established with 2-AAF/partial hepatectomy at dosages of 10～20mg/kg.Those oval cells are hepatic stem/progenitor cells expressing OV6/CK19/albumin/AFP/connexin43/C-kit.
出处
《基础医学与临床》
CSCD
北大核心
2004年第3期344-347,i002,共5页
Basic and Clinical Medicine
关键词
卵圆细胞
肝前体细胞
干细胞
免疫组化
oval cell
hepatic progenitor cell
stem cell
immunohistochemistry
