IGF2在小鼠胚胎干细胞定向分化为胰岛样细胞过程中的表达

The Expression of IGF2 in Mouse Embryonic Stem Cells to Differentiate into Islet-Like Cells

目的观察印迹基因胰岛素样生长因子(IGF)2在小鼠胚胎干细胞定向诱导分化为胰岛样细胞过程中的表达情况。方法体外诱导胚胎干细胞向胰岛样细胞分化,逆转录-PCR(RT-PCR)和细胞免疫荧光检测胰岛细胞标志基因表达;聚合酶链式反应-限制性内切酶片段长度多态性(PCR-RFLP)检测印迹基因IGF2在诱导分化前后细胞中的亲本表达情况。结果诱导分化终末细胞能表达胰岛素、胰高糖素及C肽等胰岛细胞特异性标志物,PCRRFLP检测分析显示,体外诱导分化的细胞的印迹基因IGF2呈双等位基因表达,印迹丢失。结论体外诱导培养可导致胚胎干细胞印迹基因IGF2表达的改变。

Objective To observe the effects of insulin-like growth factor-2 （IGF2） in the course of mouse embryon- ic stem cells induced to differentiate into islet-like cells. Methods Mouse ES cells were induced to differentiate into islet- like cells in vitro. The expression of islet specific markers was tested by RT-PCR and immunofluorescence assay. RT-PCR/ RFLP was used to test the imprinted genes IGF2 parental expression in cells at different stages, Results Islet specific mark- ers were expressed in differentiated cells, such as insulin, glucagon and C-peptide. PCR-RFLP showed that imprinted genes IGF2 derived from embryonic stem cells were biallelic expression and loss of imprinting. Conclusion Gene imprinting sta- tus of IGF2 was changed in differentiated cells in vitro.