摘要
目的 采用骨髓间质干细胞体外诱导为心肌细胞 ,为心力衰竭的干细胞移植治疗提供新的细胞材料。方法 抽取贵州香猪骨髓液 3ml,按照Wakitani的方法培养出骨髓间质干细胞 ,连续传代 4个月 ,得到永生型细胞 ,经 5 氮胞苷 (5 azacytidine)刺激后 ,进行RT PCR ,基因测序 ,免疫组化 ,电镜超微结构鉴定。结果 骨髓间质干细胞经 5 氮胞苷刺激后 ,部分细胞呈梭形。RT PCR ,基因测序结果表明细胞有心房利钠肽 (ANP) ,脑利钠肽 (BNP) ,心肌特异性肌球蛋白重链 (cardiac MHC) ,β肌球蛋白重链 (beta MHC) ,α骨骼肌肌动蛋白 (α skeletalactin)表达 ,结蛋白 (desmin) ,肌球蛋白重链 (MHC) ,心肌特异性肌钙蛋白Ⅰ (cTnTⅠ )免疫组化阳性。电镜示梭形细胞有明显的肌丝 ,细胞核呈单椭圆形 ,位于细胞中央。结论 骨髓间质干细胞经 5 氮胞苷刺激后 ,从基因 ,蛋白 ,超微形态上已有心肌细胞特点 ,表明在体外环境下可向心肌细胞转化 。
Objective To procure stem cells and generate into cardiomyocytes from porcine marrow mesenchymal source,providing potential therapeutic application for heart failure. Methods 3 ml bone marrow was extracted from the posterior superior iliac spine of Gui zhou Xiang porcine. Mesenchymal stem cells were cultured according to Wakitani′s methods. Immortalized cells were obtained by frequent subculture for 4 months. After being co cultured with 5 azacytidine, the cultured cells were evaluated by transmission electron microscope, immunostaining, RT PCR and gene sequence analysis. Results After being co cultured with 5 azacytidine, some of MSCs became spindle like cells. RT PCR and gene sequence analysis showed that these cells expressed ANP, BNP, cardiac MHC, β MHC, α skeletal actin. These cells were found positively stained with MHC, desmin, and cTnT Ⅰ. Transmission electron microscope showed that myofilaments were formed and oval nucleus lying in the center of the cell. Conclusion Cardiomyocytes can be generated from MSCs. It suggests that MSCs might be a new cell source for stem cells transplantation in heart failure therapy.
出处
《中华心血管病杂志》
CAS
CSCD
北大核心
2004年第1期59-63,共5页
Chinese Journal of Cardiology
基金
国家自然科学基金资助课题 (3 0 0 70 748)
