5-氮胞苷诱导脂肪间充质干细胞向心肌细胞分化的毒理遗传学研究
Toxicological genetics of the 5-azacyti-dine in inducing adipose-derived mesenchymal stem cells differentiate into myocardial cells
张卫泽 1李国庆 2陈永清 1马凌 1秦勉 1韩娟萍2
作者信息
- 1. 兰州军区兰州总医院心血管内科,甘肃兰州730050
- 2. 兰州大学第二临床医学院,甘肃兰州730000
- 折叠
摘要
目的:观察5-氮胞苷(5-aza)对成人脂肪间充质干细胞(ADMSCs)向心肌细胞方向诱导分化的毒理遗传学影响.方法:自成人脂肪组织中分离培养ADMSCs,用5-aza对ADMSGs进行诱导,设立不同诱导浓度的Ⅰ ,Ⅱ,Ⅲ,(5,10,20μmoL/L)组及对照组Ⅳ,光镜下观察细胞的生长情况及形态学变化,免疫细胞化学方法检测心肌特异性肌钙蛋白I(cT-nI),RT-PCR方法检测cTnI基因的表达,流式细胞仪检测细胞周期及凋亡,并对各诱导组常规核型分析.结果:诱导后4wk Ⅰ,Ⅱ,Ⅲ组ADMSCs呈现类似心肌细胞的形态学特征.Ⅰ,Ⅱ,Ⅲ组免疫组化结果见cTnI阳性表达,RT-PCR条带见cTnI基因表达.各组染色体结构及数目均未见异常.结论:5-aza作为诱导剂对ADMSCs遗传性无不良影响.
Abstract
AIM:To observe toxicological genetics effects of 5-azacytidine (5-aza) in inducing the adipose-derived mesenchymal stem cells ( ADMSCs) into myocardial cells. METHODS: ADMSCs were isolated from adipose tissue. The ADMSCs were induced with 5-aza in different concentrations groups(5, 10,20 (μmol/L) and control. Cells growth and morphological changed were observed with light microscope, immunocytochemistry detected cardiac-specific troponin I ( cTnl). RT-PCR was used to detect cTnI gene expression. Flow cytometry was used to detect cell cycle and apoptosis. Chromosome structure was detected with conventional karyorype analysis. RESULTS: The ADMSCs in group I, II, III presented similar myocardial morphological characteristics after being induced 4 weeks. The ADMSCs in I, Ⅱand Ⅲ groups show the cTnI positive in protein and gen levels. Chromosome structure in each groups had no abnormal chanes. CONCLUSION: As an inductor,5-aza had no harmful effects about inheritance to ADMSCs.
关键词
脂肪间充质干细胞/5-氮杂胞苷/核型/细胞遗传/毒理遗传学Key words
adipose-derived mesenchymal stem cells/5-azacyti-dine/karyotype/cytoplasmic inheritance/toxicological genetics引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据