摘要
目的:探讨抗坏血酸作为诱导剂对小鼠胚胎干细胞(mESC)分化为心肌细胞的影响,建立一种体外诱导mESC分化为心肌细胞的实验方法。方法:采用直接悬浮培养法使mESC形成拟胚体(EBs),用含不同浓度抗坏血酸的分化培养基对其进行诱导分化,摸索最佳的诱导分化条件。结果:抗坏血酸诱导mESC分化为心肌细胞的最佳浓度为0.1mg/ml,约为70%的EBs出现跳动的心肌合胞体,显著高于不添加任何诱导剂的对照组。抗坏血酸诱导产生的心肌细胞表达多种心肌蛋白,具有心肌细胞的结构特征。结论:抗坏血酸能够促进mESC向心肌细胞分化,应用抗坏血酸体外诱导mESC向心肌细胞分化是一种较为理想的体外诱导方法。
Objective： To evaluate the effect of ascorbic acid on the myocardiac differentiation of mouse embryonic stem cells （mESC） and establish a method of in vitro differentiation of mESC toward cardiomyocytes. Methods： After 5 days of suspension, embryoid bodies （EBs） formed from mESC were seeded in 24-well plates and incubated with differentiation medium containing ascorbic acid in various concentration to tincl the optimal concentration for myocardiac differentiation. The beating EBs were calculated and the beating cells were immunostained with cardiomyocyte-specific markers. RT-PCR was used to identify the genes expressed in these cells. Results： The maximum cardiac differentiation rate of mESC was reached at a concentration of 0.1 mg/ml ascorbic acid, while the beating cardiomyocytes were found in approximately 70 percentage of EBs. There was significant difference compared with the control. The mESC- derived cardiomyocytes induced by ascorbic acid were characterized by the expression of special cardiac genes and pro- teins and had the similar structures with cardiomyocytes. Conclusion： Ascorbic acid could enhance the differentiation of mESC into cardiomyocytes, and it could serve as a valuable strategy for myo cardiac differentiation of mESC in vitro.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2007年第4期392-396,共5页
Chinese Journal of Anatomy
基金
国家自然科学基金重点项目(30530220)
北京市自然科学基金项目(7062053)
关键词
抗坏血酸
胚胎
干细胞
心肌细胞
细胞分化
ascorbic acid； embryo； stem cells
cardiomyocytes； cell differentiation
