摘要
目的:体外分离和定向诱导小鼠胎肝间充质干细胞向胰岛B样细胞分化。方法:无菌条件下从正常C57BL／6J胎鼠肝中分离出间充质干细胞,体外培养传3代后用高浓度葡萄糖培养基以及碱性纤维生长因子(basic fihroblast growth factor,bFGF)和尼克酰胺诱导分化,观察胎肝间充质干细胞诱导前后形态变化;用RT-PCR检测细胞诱导前后胰十二指肠同源异型基因盒1(pancreatic duodenal homeobox-1,PDX-1)、胰岛素原1(proinsulin-1,INS-1)、葡萄糖转运子2(glucose transporter-2,GLUT-2)表达情况;胰岛素免疫细胞化学染色鉴定诱导后细胞胰岛素的表达;在形成胰岛样细胞簇后,用双硫腙做胰岛B细胞特异性染色。结果:RT-PCR显示诱导5 d后PDX-1、INS-1、GLUT-2均有表达,而诱导前的细胞则没有检测到表达;胰岛素免疫细胞化学表明细胞簇内的细胞胰岛素染色强阳性;细胞簇双硫腙染色阳性(每个T-25培养瓶有80-120个)。结论:从胎肝中分离出的间充质干细胞在体外可以定向诱导分化为胰岛B样细胞。
Objective:To isolate mesenchymal stem cells (MSCs) from mouse fetal liver and induce them differentiate into islet B-like cells. Methods:MSCs were isolated from C57BL/6J mouse fetal liver and were induced with high concentration of glucose, basic fibroblast growth factor(bFGF) ,and nicotamine medium. The gene expressions related to islet B cells such as pancreatic duodenal homeobox-1 (PDX-1), proinsulin-1 (INS-1) ,and glucose transporter-2 (GLUT-2) were detected by RT-PCR. Insulin in the treated cells was examined by immunocytochemistry. The insulin clusters were stained with dithizone (DTZ), a zinc-chelating agent known to selectively stain pancreatic B cells. Results: RT-PCR showed that the treated cells expressed PDX-1, INS-1 and GLUT-2, while the undifferentiated cells did not. After approximately 10 d of treatment, the fetal liver cells formed DTZ-stained cell clusters in flasks (80-120 clusters in a flask 25 cm2 in area). Immunocytochemistry also confirmed that these aggregates were strongly positive for insulin. Conclusion: MSCs derived from fetal liver can be induced into islet B-like cells in vitro.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2004年第8期827-830,共4页
Academic Journal of Second Military Medical University
关键词
间充质干细胞
胎肝
胰岛B样细胞
细胞分化
mesenchymal stem cells
fetal liver
islet B-like cells
cell differentiation
