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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 explore the possibility of human umbilical cord mesenchymal stem cells (UC-MSCs) differentiating into insulin-secreting cells. Methods UC-MSCs were isolated from umbilical cord by direct adherent method. The third generation of cells were identified by flow cytometry. UC-MSCs (induction group) were induced with β-mercaptoethanol, nicotinamide and basic fibroblast growth factor (bFGF), and the control group was cultured in a medium without any inducing agent. The morphological changes of UC-MSCs were observed under an inverted microscope. Cells in the three weeks after induction were stained with dithizone. The levels of insulin in the culture supernatant were determined by chemiluminescence immunoassay. The expression of related genes in islet cells was detected by RT-PCR. As a result, UC-MSCs-associated antigens CD90, CD105 and CD13 were highly expressed, whereas CD34, CD45 and HLA-DR were negatively expressed. The morphology of UC-MSCs was changed from spindle to round or oval shape after induction, and dithizone staining was positive. 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 cells expressed islet cell-related genes after induction. Conclusion UC-MSCs have the potential to differentiate into insulin-secreting cells.