小鼠胚胎干细胞定向诱导为胰岛样细胞过程中印记基因变化

Expression of imprinted genes during the course of differentiation from mouse embryonic stem cells to islet-like cells in vitro

摘要:

目的：探讨体外诱导对小鼠胚胎干细胞株SF1-G印记基因Kcnq1、Cdkn1c的影响。方法：体外诱导小鼠胚胎干细胞SF1-G向胰岛样细胞分化，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)检测印记基因Kcnq1、Cdkn1c在分化各阶段细胞中的亲本表达情况。结果：胰岛细胞标志性基因Insulin、Glucagon、Somatostatin、IAPP、Glut2在诱导分化后的细胞都有表达；分化终末细胞能表达胰岛细胞特异性蛋白胰岛素、C肽及Somastatin；PCR-RFLP检测分析显示，体外诱导分化后细胞的印记基因Kcnq1、Cdkn1c呈双等位基因表达。结论：胚胎干细胞经体外诱导培养可定向分化为胰岛样细胞；用此种分化方法可导致胚胎干细胞印记基因表达的改变，出现印记丢失。［中国当代儿科杂志，2010，12(12)：954-958］

Abstract:

OBJECTIVE: To study the effects of in vitro inducement on the expression of SF1-G imprinted genes, Kcnq1 and Cdkn1c during the course of differentiation from mouse embryonic stem (ES) cells to 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 or immunofluorescence. RT-PCR/RFLP was used to test the imprinted genes parental expression in cells at different stages. RESULTS: Islet specific genes, such as Insulin, Glucagon, Somatostatin, IAPP and Glut2, were expressed in differentiated cells. The proteins of insulin, C-peptide and Somastatin were expressed in the final stage cells. Imprinted gene Kcnq1 and Cdkn1c were biallelicly expressed in islet-like cells. CONCLUSIONS: Mouse ES cells can be successfully induced into islet-like cells in vitro. Gene imprinting status of Kcnq1 and Cdkn1c may be changed in differentiated cells (causing loss of imprinting) during the in vitro inducement.［Chin J Contemp Pediatr, 2010, 12 (12):954-958］