拟胚体贴壁时间对小鼠胚胎干细胞心肌分化的影响

发布时间：2018-03-29 11:02

本文选题：胚胎干细胞　切入点：心肌细胞　出处：《基础医学与临床》2013年09期

【摘要】：目的观察拟胚体(EB)贴壁时间对小鼠胚胎干细胞(ESC)心肌分化的影响并研究其机制。方法用悬滴培养法促进ESCs形成EBs。EBs在不同的分化天数贴壁,观察搏动EBs百分比,RT-PCR检测Nkx2.5,GATA4和β-MHC mRNA表达,Western blot检测Src家族酪氨酸激酶磷酸化水平。结果分化第3,4天贴壁组搏动EB百分比及Nkx2.5,GATA4,β-MHC表达水平显著低于分化第5,6和7天贴壁组(P0.05);EB贴壁能够使Src激酶磷酸化水平升高,Src激酶阻断剂PP2能够抑制贴壁诱发的Src激酶磷酸化水平升高(P0.05);对于分化第4天贴壁组,在分化第4～6天使用PP2能够增加搏动EBs百分比及β-MHC表达水平(P0.05)。结论 EB贴壁时间是影响ESC心肌分化的一个重要因素。在分化第4天或者之前贴壁可以显著抑制心肌分化,其机制可能是EB贴壁激活了Src激酶。
[Abstract]:Objective to observe the effect of adherent time of EBS on myocardial differentiation of mouse embryonic stem cell ESCs and its mechanism. Methods suspension culture method was used to promote the formation of ESCs into EBs.EBs and adhere to the wall at different differentiation days. The expression of GATA4 and 尾 -MHC mRNA was detected by RT-PCR. The tyrosine kinase phosphorylation of Src family was detected by Western blot. Results the percentage of EB pulsatile and the expression of Nkx2.5, GATA4, 尾 -MHC in adherent group were significantly lower than those in 5th and 7th day of differentiation. P0.05 EB adherent can increase the level of phosphorylation of Src kinase. PP2, a blocker of SRC-kinase, can inhibit the increase of Src kinase phosphorylation induced by adhesion. The percentage of pulsatile EBs and the expression level of 尾 -MHC were increased by using PP2 on the 4th day of differentiation. Conclusion EB adhesion time is an important factor affecting myocardial differentiation of ESC. Adhesion on or before the 4th day of differentiation can significantly inhibit myocardial differentiation. The mechanism may be that EB adherent activates Src kinase.
【作者单位】：武汉大学中南医院心内科;南方医科大学神经生物学教研室;
【基金】：国家自然科学基金(81200119) 中央高校基本科研业务费专项资金(121007)
【分类号】：R329.1

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