小鼠骨髓基质干细胞体外分化为前体心肌细胞的初步实验

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (02): 190-194.

小鼠骨髓基质干细胞体外分化为前体心肌细胞的初步实验

张勇¹, 蔡振杰², 陈如坤³

1. 济南军区总医院心外科, 山东, 济南, 250031;
2. 第四军医大学西京医院心血管外科, 陕西, 西安, 710033;
3. 浙江大学医学院第二附属医院, 浙江, 杭州, 310009

出版日期:2005-02-20 发布日期:2005-02-20
基金资助:
收稿日期:2004-10-25。
作者简介:张勇(1972- ),男,主治医师,在读博士研究生,E-mail:zhywhzh@163.net

Experimental study of differentiation of mouse bone marrow stromal stem cells into progenitor cardiomyocyte in vitro

ZHANG Yong¹, CAI Zhen-jie², CHEN Ru-kun³

1. 济南军区总医院心外科, 山东, 济南, 250031;
2. 第四军医大学西京医院心血管外科, 陕西, 西安, 710033;
3. 浙江大学医学院第二附属医院, 浙江, 杭州, 310009

Online:2005-02-20 Published:2005-02-20

摘要/Abstract

摘要： 目的研究体外诱导小鼠骨髓基质干细胞(MSCs)向心肌细胞分化的可能性。方法 MSCs通过细胞传代培养,并通过流式细胞仪进行鉴定。MSCs经过5-杂氮胞苷诱导后,通过RT-PCR、半定量RT-PCR、Western-blotting和透射电镜检测诱导后的MSCs。结果培养的MSCs为形态均一的梭形细胞,有时可见细胞融合。流式细胞仪显示诱导后的MSCsCD29、CD44表达阳性,而CD34、CD45表达阴性。经过5-杂氮胞苷诱导后的MSCs表达Nkx2-5/Csx、GATA4、β-MHC基因和α-sarcomericactin和desmin蛋白,透射电镜显示诱导后的MSCs有肌丝形成。诱导后的MSCsDNA甲基化转移酶表达降低。结论小鼠MSCs能够向前体心肌细胞分化。

Abstract: Objective To investigate the possibility of inducing mouse bone marrow stromal stem cells(MSCs) into progenitor cardiomyocytes in vitro. Methods The MSCs were isolated by adhesion culture in vitro and flow cytometery was employed to identify the phenotypes of the cell passages. 5-azacytidine was used to induce the stem cells to differentiate into cardiomyotes in the experimental group, which were then detected by RT-PCR, semi-quantitative RT-PCR, Western-blot analysis, electron microscopy and immunofluorescence technique. Results The isolated subcultured MSCs displayed a fusiform cell-like morphology. The MSCs were uniformly positive for CD29 and CD44 but negative for CD34 and CD45, and after induction, they expressed cardiomyocyte-specific transcription factors(NKx2-5/Csx and GATA4) and fetal ventricular cardiomyocyte-specific gene β-myosin heavy chain, but not adult ventricular cardiomyocyte-specific gene α-myosin heavy chain as detected by RT-PCR. Western blotting identified the expressions of α-sarcomeric actin and desmin in the induced MSCs, with myofilament formation observed under electron microscope. Compared with the control group, the expression level of DNA methyltransferase mRNA was significantly lowered in the experimental group as observed by semi-quantitative RT-PCR(P<0.05). Conclusion MSCs are capable of differentiating into progenitor cardiomyocytes.

中图分类号:

R329.24

张勇¹, 蔡振杰², 陈如坤³. 小鼠骨髓基质干细胞体外分化为前体心肌细胞的初步实验[J]. 南方医科大学学报, 2005, 25(02): 190-194.

ZHANG Yong¹, CAI Zhen-jie², CHEN Ru-kun³. Experimental study of differentiation of mouse bone marrow stromal stem cells into progenitor cardiomyocyte in vitro[J]. Journal of Southern Medical University, 2005, 25(02): 190-194.

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