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-azacytidine in inducing adipose-derived mesenchymal stem cells differentiate into myocardial cells

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ADMSCs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.

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（cTnI）.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 karyotype analysis.RESULTS：The ADMSCs in group I,II,III presented similar myocardial morphological characteristics after being induced 4 weeks.The ADMSCs in I,II and 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.