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|本期目录/Table of Contents|

5-氮胞苷体外诱导骨髓间充质干细胞向心肌样细胞的分化

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2009年第6期
页码:
801-804
栏目:
基础研究
出版日期:
2009-11-05

文章信息/Info

Title:
Differentiation of human mesenchymal stem cells into cardiomyocyte-like cells in vitro induced by 5-azacytidine
作者:
曹京燕1; 程月新2; 姜敏辉3; 胡桂萍1; 王卫彪3; 姜胜华4
南通大学：第四附属医院:1.心内科，2.血液科，江苏 盐城224006， 第一附属医院:3.心内科，4.血液科，江苏 南通226001
Author(s):
CAO Jing-yan1;  CHENG Yue-xin2;  JIANG Min-hui3;  HU Gui-ping1;  WANG Wei-biao3;  JIANG Sheng-Hua4
1.Department of Cardiology, 2.Department of Hematology, Fourth Affiliated Hospital, Yancheng 224006, Jiangsu, China, 3.Department of Cardiology, 4.Department of Hematology, First Affiliated Hospital, Nantong University, Nantong 226001, Jiangsu, China
关键词:
骨髓间充质干细胞; 5-氮胞苷; 细胞分化; 心肌样细胞
Keywords:
bone marrow mesenchymal stem cells;  5-azacytidine;  cellular differentiation;  cardiomyocyte-like cells
分类号:
R329.21
DOI:
-
文献标识码:
A
摘要:
目的: 研究5-氮胞苷（5-Aza）对培养人骨髓间充质干细胞（MSC）的作用，并对分化后的心肌样细胞进行鉴定。方法: 采用密度梯度离心法分离到骨髓单个核细胞（MB-MNC），用含200 ml/L胎牛血清的低糖型DMEM培养液进行培养。采用差速贴壁法纯化MSC，用流式细胞仪检测细胞表面抗原。以5-Aza诱导第3代MSC 24 h后继续培养。培养4周，用免疫细胞化学染色法检测肌系标记抗原:α-肌动蛋白（α-actin）及心肌细胞特异性标记抗原：肌钙蛋白T（cTnT）；在透射电镜下观察细胞的超微结构。结果: MSC经5-Aza诱导分化后，可表达α-actin和cTnT，未经诱导的同培养天数的MSC中均未见表达。透射电镜可观察到肌丝等心肌细胞的特异性结构。结论: 5-Aza可诱导MSC分化为心肌样细胞。
Abstract:
AIM: To observe the effect of 5-azacytidine (5-Aza) on bone marrow mesenchymal stem cells (MSCs) and to determine the cardiomyocyte-like cells that were differentiated from MSCs. METHODS: Bone marrow mononuclear cells (BM-MNC) were isolated from bone marrow by the method of density gradient centrifugation and were cultured in low glucose DMEM containing 200 ml/L fetal bovine serum. The cells were purified by the different abilities of adhesion. Cell surface antigens were tested by flow cytometry. MSCs were induced by 5-Aza at the third passage for 24 h. After being treated by 5-Aza for 4 weeks, the cells were harvested. Muscle tissue antigen such as α-actin and the cardiac muscle heterologous protein, e.g., cTnT, were detected by immunocytochemical staining. Ultramicrostructure of the cells was observed under transmission electron microscope. RESULTS: After being induced by 5-Aza, α-actin and cTnT were expressed in MSCs. However, expression in MSC without 5-Aza treatment was negative. Ultramicrostructure of the cells such as myofilament was observed under transmission electron microscope. CONCLUSION: Human bone marrow MSCs can differentiate into cardiomyocyte-like cells under 5-Aza induction in vitro.

参考文献/References

［1］ Tsai RY, Kittappa R, McKayRD. Plasticity, niches, andthe use of stem cells［J］. Dev Cell, 2002, 2(6):707-712.

［2］ Bosnakovski D, Mizuno M, Kim G, et al. Isolation and multilineag differentiation of bovine bone marrow mesenchymal stem cells［J］. Cell Tissue Res, 2005, 319(2):243-253.

［3］ Gronthos S, Zannettino AC, Hay SJ, et al. Molecular and cellular characterization of highly purified stormal stem cells derived from human bone marrow［J］. J Cell Sci, 2003, 116(Pt 9):1827-1835.

［4］ Fukuda K. Development of regenerative cardiomyocytes from msenchy-mal stemcells for cardiovascular tissue engineering［J］. Artif Organs, 2001, 25(3):187-193.

［5］ Hwang JH, Yuk SH, Lee JH, et al. Isolation of muscle derived stem cells from rat and its smooth muscle differentiation［J］. Mol Cells, 2004, 17(1):57-61.

［6］ Panteghini M. Present issues in the determination of troponins and other makers of cardiac damage［J］. Clin Biochem, 2000, 33(3):161-166.

备注/Memo

备注/Memo:
收稿日期：2008-7-31.基金项目:江苏省盐城市医学重点人才资助(2007013)；盐城市科技计划项目资助(2007045) 通讯作者：姜敏辉,副教授,主要从事冠心病基础与临床研究Email：jiangminhui@medmail.com.cn. 作者简介：曹京燕， 副主任医师，硕士Email：yccyx68@yahoo.com.cn
更新日期/Last Update: 2009-09-30