体外定向诱导人脐带间充质干细胞分化为胰岛样细胞

背景：人脐带间充质干细胞具有获取容易、免疫原性低等优点，目前尚缺乏体外诱导其分化为胰岛样细胞的成熟方案。目的：探讨人脐带间充质干细胞在体外一定条件下分化为胰岛样细胞的可能性。方法：无菌条件下分离培养人脐带间充质干细胞，细胞传3代后，采用两步法诱导其向胰岛样细胞分化：第1步，加入含100pg／L13-神经生长因子，4nmol／LActivinA，10mmol/L尼克酰胺，25pg／L表皮生长因子，体积分数为10％胎牛血清的DMEM／F12培养基中培养7d；第2步，将诱导液换为含10mmoI／L尼克酰胺，10pg／L碱性成纤维细胞生长因子，1％胰岛素一转铁蛋白一硒的DMEM／F12培养基，诱导时间为14d。对照组单纯加入DMEM／F12培养基进行培养。结果与结论：诱导2周后脐带间充质干细胞开始聚集成团，诱导3周后，葡萄糖刺激试验阳性、PDX-1和Insulin基因表达。而对照组细胞无胰岛素分泌，胰岛细胞相关基因表达阴性。实验成功地将脐带间充质干细胞诱导成了胰岛样细胞。

Oriented differentiation of human umbilical cord mesenchymal stem cells into islet-like cells in vitro

BACKGROUND： Human umbilical cord mesenchymal stem cells have some advantages such as easy acquirement and low immunity. But there is no mature program for differentiating into islet-like cells in vitro at present. OBJECTIVE： To investigate the possibility of human umbilical cord mesenchymal stem cells differentiating into islet-like cells in appropriate conditions. METHODS： The human umbilical cord mesenchymal stem cells were isolated and cultured under sterile conditions, following passage for three generations. The human umbilical cord mesenchymal stem cells were induced to differentiate into islet-like cells by two steps. Step 1： the cells were cultured in the dulbecco＇s modified eagle＇s medium/F12 culture medium containing 100 pg/L 13-nerve growth factors, 4 nmol/L activinA, 10 mmol/L nicotinamide, 25 pg/L epidermal growth factor and fetal bovine serum with the volume fraction of 10% for 7 days. Step 2： the induction medium was changed into the dulbecco＇s modifiedeagle＇s medium/F12 culture medium containing 10 mmol/L nicotinamide, 10 IJg/L alkaline fibroblast growth factors and 1% insulin-transferdn-selenium, and induced for 14 days. The cells in the control group were cultured with the dulbecco＇s modified eagle＇s medium/F12 culture medium. RESULTS AND CONCLUSION： Cells began to aggregate after induced for 2 weeks. After 3 weeks of induction, the glucose stimulation test was positive with the expression of pancreatic and duodenal homeobox 1 and insulin No insulin secretion was found in the control group and the expression of islet cells related gene was negative in the control group. The human umbilical cord mesenchymal stem cells were successfully induced to differentiate into the islet-like cells.