摘要
目的探讨人脐带间充质干细胞(UC-MSCs)向胰岛素分泌细胞分化的可能性。方法采用直接贴壁法从脐带中分离UC-MSCs,取第3代细胞作流式细胞术鉴定其表型。用β-巯基乙醇、尼克酰胺和碱性成纤维生长因子(bFGF)诱导UC-MSCs(诱导组),对照组在未添加任何诱导试剂的培养基中培养。倒置显微镜下观察UC-MSCs的形态学变化;取诱导后3周的细胞进行双硫腙染色,采用化学发光免疫法测定培养上清液中胰岛素水平,RT-PCR检测胰岛细胞相关基因的表达。结果细胞高表达UC-MSCs相关抗原CD90、CD105和CD13,而低表达造血细胞相关抗原CD34、CD45和HLA-DR。UC-MSCs经诱导后细胞形态由梭形变为圆形或椭圆形,双硫腙染色为阳性。诱导组胰岛素水平明显高于对照组[(0.305±0.065)μU/ml vs.(0.085±0.024)μU/ml](P<0.01)。RT-PCR显示诱导后细胞表达胰岛细胞相关基因。结论 UC-MSCs具有向胰岛素分泌细胞分化的潜能。
Objective To investigate the feasibility of human umbilical cord mesenchymal stem cells（UC-MSCs） differentiating towards insulin-secreting cells in vitro. Methods UC-MSCs were isolated from umbilical cord by directly adherence growth, and the surface antigens in the third generation of UC-MSCs were identified by flow cytometry. The cells were treated with mercaptoethanol,nicotinamide and bFGF in induction group and those were not treated as control group. The morphologic changes of UC-MSCs were observed under inverse microscope, and the cells induced for 3 weeks were stained with dithizone. The level of insulin in the cultured supernatant was detected by chemiluminescence immunoassay, and the expressions of islet cell-related genes were measured by RT-PCR. Results UC-MSCs-related antigens CD90, CD105 and CD13 were highly expressed,while hematopoietic cells-related antigens CD34,CD45 and HLA-DR were lowly expressed in UC-MSCs. UC-MSCs induced in the differentiation medium were gradually transformed into round or oval in shape with positive dithizone staining. The level of insulin in induction group was significantly higher than that in control group[（0. 305±0. 065） μU/ml vs. （0. 085±0. 024） μU/ml] （P〈0. 01）. RT-PCR showed that the induced cells also expressed islet cell-related genes. Conclusion UC-MSCs are able to differentiate towards insulin-producing cells.
出处
《江苏医药》
CAS
北大核心
2013年第9期1021-1024,F0002,共5页
Jiangsu Medical Journal
基金
南京市医学科技发展项目(ykk10090)
关键词
脐带间充质干细胞
胰岛素分泌细胞
Umbilical cord mesenchymal stem cell
Insulin-secreting cells
