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5-氮杂胞苷诱导持续时间对人孕早期胎儿间充质干细胞向心肌样细胞分化的影响。

Effect of 5-azacytidine induction duration on differentiation of human first-trimester fetal mesenchymal stem cells towards cardiomyocyte-like cells.

作者信息

Zhang Yihua, Chu Yuankui, Shen Wenzheng, Dou Zhongying

机构信息

Shaanxi Branch of National Stem Cell Engineering and Technology Centre, College of Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.

出版信息

Interact Cardiovasc Thorac Surg. 2009 Dec;9(6):943-6. doi: 10.1510/icvts.2009.211490. Epub 2009 Sep 28.

DOI:10.1510/icvts.2009.211490
PMID:19786397
Abstract

The aim of this study is to investigate effects of 5-azacytidine (5-aza) induction duration on differentiation of bone marrow mesenchymal stem cells (MSCs) from human first-trimester abortus (hfMSCs) towards cardiomyocyte-like cells. hfMSCs were stimulated with 10 micromol/l 5-aza for 24 h (group A), 48 h (group B) and 21 days (group C), respectively. During the induction, 30-40% of the cells gradually enlarged, elongated, connected with adjoining cells and formed myotube-like structures, branches and string-bead-like nuclei. Some of the cells congregated into cell clusters or strips. After the induction, numerous myofilaments in the cytoplasm and conjunction of intercalated disc-like structure between adjoining cells were observed. The induced cells expressed messenger ribonucleic acids (mRNAs) and proteins of myocardium-specific alpha-actin, sarcomeric beta-myocin heavy chain and troponin-T. The positive cell percentages for the three antigens in group C were each significantly higher than those antigens in group A and B (P<0.01) and the cell population doubling time (PDT) of group C was longer than those of group A and B (P<0.01). These indicate that 21-d induction with 10 micromol/l 5-aza slows down proliferation speed of hfMSCs but increases differentiation rate of hfMSCs into cardiomyocyte-like cells if compared with 24-48 h induction.

摘要

本研究旨在探讨5-氮杂胞苷(5-aza)诱导持续时间对人早孕流产胎儿骨髓间充质干细胞(hfMSCs)向心肌样细胞分化的影响。分别用10微摩尔/升的5-aza刺激hfMSCs 24小时(A组)、48小时(B组)和21天(C组)。诱导过程中,30%-40%的细胞逐渐增大、伸长,与相邻细胞相连,形成肌管样结构、分支和串珠样核。部分细胞聚集成细胞簇或条带。诱导后,观察到细胞质中有大量肌丝,相邻细胞间有闰盘样结构连接。诱导细胞表达心肌特异性α-肌动蛋白、肌节β-肌球蛋白重链和肌钙蛋白-T的信使核糖核酸(mRNA)和蛋白质。C组三种抗原的阳性细胞百分比均显著高于A组和B组(P<0.01),且C组细胞群体倍增时间(PDT)长于A组和B组(P<0.01)。这些表明,与24-48小时诱导相比,用10微摩尔/升的5-aza诱导21天会减慢hfMSCs的增殖速度,但会提高hfMSCs向心肌样细胞的分化率。

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