天津医药

• 实验研究 •    下一篇

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

刘峰   

  1. 南昌大学第一附属医院内分泌科
  • 收稿日期:2013-04-01 修回日期:2013-06-15 出版日期:2013-11-15 发布日期:2013-11-15
  • 通讯作者: 刘峰

The Expression of IGF2in Mouse Embryonic Stem Cells to Differentiate into Islet-Like Cells

LIU Feng   

  1. Department of Endocrinology, The Affiliated Hospital of Nanchang University
  • Received:2013-04-01 Revised:2013-06-15 Published:2013-11-15 Online:2013-11-15
  • Contact: LIU Feng

PDF (PC)

2063

摘要：

【摘要】  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。

关键词: 胚胎干细胞, 基因组印迹, 胰岛素样生长因子Ⅱ, 等位基因, 细胞分化

Abstract:

[Abstract]   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 isletlike 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 IGF2parental expression in cells at different stages.   Results  Islet specific markers were expressed in differentiated cells, such as insulin, glucagon and C-peptide. PCR-RFLP showed that imprinted genesIGF2derived from embryonic stem cells were biallelic expression and loss of  imprinting . Conclusion   Gene imprinting status of IGF2was changed in differentiated cells in vitro.

Key words: embryonic stem cell, genomic imprinting, insulin-like growth factor Ⅱ, alleles, cell differentiation