探讨大鼠脂肪间充质干细胞（adipose-derived mesenchymal stem cells，AMSC）体外分化成心肌样细胞的潜能，为自体干细胞移植治疗心肌梗死提供理论基础.采用消化法分离大鼠AMSC，培养于RPMI1640生长培养基中，倒置相差显微镜观察细胞形态发现，随着培养时间的延长，细胞形态趋向于心肌细胞，SQ RT-PCR检测表达心肌特异性基因：β-肌球蛋白重链（β-MHC）、α-肌球蛋白重链（α-MHC）、心房利钠肽（ANP）、心肌肌钙蛋白（cTnT）、心肌肌动蛋白、肌肉增强因子和GATA-4；免疫细胞化学和免疫荧光染色检测表达心肌细胞特异性蛋白：结蛋白、横纹肌辅肌动蛋白、心肌肌动蛋白和间隙连接蛋白45(connexin 45)；Western印迹检测表达心肌特异性蛋白Nkx2.5. 实验表明，大鼠AMSC在体外培养条件下能分化成心肌样细胞，在组织工程学及干细胞移植领域有着良好的应用前景.
Abstract
The potential to differentiate into cardiomyocyte from rat adipose-derived mesenchymal stem cells(AMSCs) was investigated, AMSCs were isolated from SD rat adipose tissue by digestion and maintained in the RPMI1640 medium. AMSCs transformed into cardiomyocyte-like cells during the long-term culture. The expression of β-MHC, ANP, cTnT, GATA-4, α-cardiac actin and MEF2C were detected by Quantitative RT-PCR. Cardiac-specific proteins, such as desmin, sarcomeric, α-actinin, α-cardiac actin and connexin 45 were detected by immuocytochemistry or immunofluorescence staining. The expression of early cardiac-specific protein Nkx2.5 were detected by Western blot. The results showed that rat AMDCs was able to differentiate into a cardiomyocyte in vitro, which could be used as a source for tissue engineering or stem cell transplantation for the treatment of ischemic myocardial infarction.
关键词
脂肪间充质干细胞（AMSC） /
心肌样细胞 /
自发分化
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Key words
adipose-derived stem cells(AMSCs) /
cardiomyocyte /
differentiation
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中图分类号：
Q254
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参考文献
［1］Zak R. Development and proliferative capacity of cardiac muscle cells［J］. Circ Res, 1974, 35(2): 17-26
［2］Stamm C, Kleine H D, Westphal B, et al. CABG and bone marrow stem cell transplantation after myocardial infarction［J］. Thorac Cardiovasc Surg, 2004，52(3):152-158
［3］Dimarakis I, Habib N A, Gordon M Y. Adult bone marrow-derived stem cells and the injured heart:just the beginning?［J］. Eur J Cardiothorac Surg, 2005，28(5):665-676
［4］Memon I A, Sawa Y, Miyagawa S, et al. Combined autologous cellular cardiomyoplasty with skeletal myoblasts and bone marrow cells in canine hearts for ischemic cardiomyopathy［J］. J Thorac Cardiovasc Surg, 2005，130(3):646-653
［5］Alvarez-Dolado M, Pardal R, Garcia-Verdugo J M, et al. Fusion of bone marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes［J］. Nature, 2003，425(6961):968-973
［6］Balsam L B, Wagers A J, Christensen J L, et al. Haematopoietic stem cells adopt mature haematopoietic fates in ischaemic myocardium［J］. Nature, 2004，428 (6983): 668-673
［7］Murry C E, Soonpaa M H, Reinecke H, et al. Haematopoietic stem cells do not transdifferentiate into cardiac myocytes in myocardial infarcts［J］. Nature, 2004，428 (6983): 664-668
［8］Sch-ffler A, Büchler C. Concise review: Adipose tissue-derived stromal cells-basic and clinical implications for novel cell-based therapies［J］. Stem Cells, 2007，25 (4):818-827
［9］Rangappa S, Fen C, Lee E H, et al.Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes［J］. Ann Thorac Surg, 2003，75(3): 775-779
［10］Yamada Y, Yokoyama S, Wang X D, et al. Cardiac stem cells in brown adipose tissue express CD133 and induce bone marrow nonhematopoietic cells to differentiate into cardiomyocytes［J］. Stem cells, 2007， 25(5): 1326-1333
［11］Yamada Y, Wang X D, Yokoyama S, et al. Cardiac progenitor cells in brown adipose tissue repaired damaged myocardium［J］. Biochem Biophys Res Commun, 2006，342(2):662-670
［12］Gaustad K G, Boquest A C, Anderson B E, et al. Diferentiation of human adipose tissue stem cells using extracts of rat cardiomyocytes［J］. Biochem Biophys Res Commun, 2004，314(2):420-427
［13］Planat-Bénard V, Menard C, André M, et al. Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells［J］. Circ Res, 2004, 94(2): 223-229
［14］Strem B M,Zhu M,Alfonso Z, et al. Expression of cardiomyocytic markers on adipose tissuederived cells in a murine model of acute myocardial injury［J］. Cytotherapy, 2005，7(3):282-291
［15］赵丽丽, 屈长青, 张国华, 等. 5-氮胞苷诱导大鼠脂肪间充质干细胞分化成肌细胞［J］. 第四军医大学学报(Zhao Li-Li, Qu Chang-Qing, Zhang Guo-Hua, et al. 5-Aza-induced differentiation of rat adipose tissue derived mesenchymal stem cells into myoblasts［J］. J Fourth Mil Med Univ), 2007，28 (22):2037-2040
［16］张国华, 屈长青, 杨公社. 全反式维甲酸抑制猪脂肪间充质干细胞的成脂分化［J］. 中国生物化学与分子生物学报 (Zhang Guo-Hua,Qu Chang-Qing, Yang Gong-She. All Trans-retinoic acid inhibit adipogenesis differentiation of porcine adipose mesenchymal stem cells［J］. Chin J Biochem Mol Biol), 2007，23 (6):474-479
［17］Song Y H, Gehmert S, Sadat S, et al. VEGF is critical for spontaneous differentiation of stem cells into cardiomyocytes［J］. Biochem Biophys Res Commun, 2007，354(4):999-1003
［18］Fukuda K, Yuasa S. Stem cells as a source of regenerative cardiomyocytes［J］. Cir Res, 2006， 98(8): 1002-1013
［19］华进联, 徐小明, 窦忠英.人类胚胎生殖细胞体外分化为心肌细胞的研究［J］.生物医学工程学杂志 (Hua Jin-Lian, Xu Xiao-Ming, Dou Zhong-Ying. Rhythmic beating cardiomyocytes derived from human embryonic germ(EG) cells in vitro［J］. J Biomed Eng), 2006，23(5):1080-1085
［20］Mauritz C, Schwanke K, Reppel M, et al. Generation of functional murine cardiac myocytes from induced pluripotent stem cells［J］. Circulation, 2008，118(5):507-517
[21[Narazaki G, Uosaki H, Teranishi M,et al.Directed and systematic differentiation of cardiovAMSCsular cells from mouse induced pluripotent stem cells［J］. Circulation, 2008，118(5):498-506
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