Expression of Myocardial Specificity Markers MEF-2C and Cx43 in Rat Bone Marrow-derived Mesenchymal Stem Cells Induced by Electrical Stimulation <i>In Vitro</i>_Journal of Biomedical Engineering

Authors：

TANGMin ¹ , YANGGang ² , JIANGJian ³ , HEXueling ¹ , LIHuiming ¹ , ZHANGMengying ¹ , WUWenchao ⁴ , LIUXiaojing ⁴ ,  LILiang ^1,4

1. Institute of Biomedical Engineering, West China College of Pre-clinical and Forensic Medicine, Sichuan University, Chengdu 610041, China;
2. Institute of Medical Informatics Engineering, College of Electrical Engineering & Information, Sichuan University, Chengdu 610065, China;
3. Department of Cardiology, West China Hospital, Sichuan University, Chengdu 610041, China;
4. Laboratory of Cardiovascular Diseases, West China Hospital, Sichuan University, Chengdu 610041, China;

Corresponding author：

LILiang, Email: LiLianghx@163.com

Keywords：

rat bone marrow-derived mesenchymal stem cells; different electrical stimulation time; myocyte-specific enhancer factor 2C; connexin 43

DOI：

10.7507/1001-5515.20150114

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Bone marrow-derived mesenchymal stem cells (BMSCs) for repairing damaged heart tissue are a new kind of important treatment options because of their potential to differentiate into cardiomyocytes. We in this experiment investigated the effect of different electrical stimulation time on the expression of myocardial specificity gene and protein in rat bone marrow mesenchymal stem cells (rBMSCs) in vitro. The rBMSCs of second or third generation were randomly divided into three groups, i.e. electrical stimulation (ES) group, 5-Azacytidine (5-Aza) group and the control group. The rBMSCs in the ES groups with complete medium were exposed to 2 V, 2 Hz, 5 ms electrical stimulation for 0.5 h, 2 h, 4 h, and 6 h respectively every day for 10 days. Those in the 5-Aza group were induced by 5-Aza (10 μmol/L) for 24 h, and then cultured with complete medium for 10 days. Those in the control group were only cultured with complete medium, without any treatment, for 10 days. The rBMSCs' morphological feature in each group was observed with inverted phase microscope. The mRNA expression of myocyte-specific enhancer factor 2C (MEF-2C) and connexin 43 (Cx43) were examined with Real-Time quantitative PCR and the protein expression of MEF-2C, Cx43 were detected with Western Blot method. The results showed that the mRNA expression level of the MEF-2C, Cx43 and the protein expression level of MEF-2C, Cx43 were significantly higher in the ES group and 5-Aza group than those in the relative control group (P < 0.05). It suggests that electrical stimulation could play a part of role in the induction of the rBMSCs to differentiate into the cariomyocyte-like cells in vitro and the effectiveness of the electrical stimulation with 2 h/d had the best in our experiement. But the mechanism how electrical stimulation promotes the differentiation of rBMSC into cardiomyocyte is still unclear.

Citation： TANGMin, YANGGang, JIANGJian, HEXueling, LIHuiming, ZHANGMengying, WUWenchao, LIUXiaojing, LILiang. Expression of Myocardial Specificity Markers MEF-2C and Cx43 in Rat Bone Marrow-derived Mesenchymal Stem Cells Induced by Electrical Stimulation In Vitro. Journal of Biomedical Engineering, 2015, 32(3): 629-634. doi: 10.7507/1001-5515.20150114 Copy

1.	BARRY F P, MURPHY J M. Mesenchymal stem cells:clinical applications and biological characterization[J]. Int J Biochem Cell Biol, 2004, 36(4):568-584.
2.	OHNISHI S, OHGUSHI H, KITAMURA S, et al. Mesenchymal stem cells for the treatment of heart failure[J]. Int J Hematol, 2007, 86(1):17-21.
3.	ANTONITSIS P, IOANNIDOU-PAPAGIANNAKI E, KAIDOGLOU A, et al. In vitro cardiomyogenic differentiation of adult human bone marrow mesenchymal stem cells. The role of 5-azacytidine[J]. Interact Cardiovasc Thorac Surg, 2007, 6(5):593-597.
4.	RADISIC M, PARK H, SHING H, et al. Functional assembly of engineered myocardium by electrical stimulation of cardiac myocytes cultured on scaffolds[J]. Proc Natl Acad Sci U S A, 2004, 101(52):18129-18134.
5.	严中琴, 杨刚, 崔燎, 等.电刺激促骨髓间充质干细胞向心肌样细胞分化[J].生物医学工程学杂志, 2013, 30(3):556-561.
6.	MINGUELL J J, ERICES A, CONGET P. Mesenchymal stem cells[J]. Exp Biol Med (Maywood), 2001, 226(6):507-520.
7.	NAGAYA N, FUJⅡT, IWASE T, et al. Intravenous administration of mesenchymal stem cells improves cardiac function in rats with acute myocardial infarction through angiogenesis and myogenesis[J]. Am J Physiol Heart Circ Physiol, 2004, 287(6):H2670-H2676.
8.	REBELATTO C K, AGUIAR A M, SENEGAGLIA A C, et al. Expression of cardiac function genes in adult stem cells is increased by treatment with nitric oxide agents[J]. Biochem Biophys Res Commun, 2009, 378(3):456-461.
9.	WANG Tingzhong, XU Zhengyun, JIANG Wenhui, et al. Cell-to-cell contact induces mesenchymal stem cell to differentiate into cardiomyocyte and smooth muscle cell[J]. Int J Cardiol, 2006, 109(1):74-81.
10.	BURLACU A, ROSCA A M, MANIU H, et al. Promoting effect of 5-azacytidine on the myogenic differentiation of bone marrow stromal cells[J]. Eur J Cell Biol, 2008, 87(3):173-184.
11.	MARUTHA RAVINDRAN C R, TICKU M K. Effect of 5-azacytidine on the methylation aspects of NMDA receptor NR2B gene in the cultured cortical neurons of mice[J]. Neurochem Res, 2009, 34(2):342-350.
12.	GENOVESE J A, SPADACCIO C, LANGER J, et al. Electrostimulation induces cardiomyocyte predifferentiation of fibroblasts[J]. Biochem Biophys Res Commun, 2008, 370(3):450-455.
13.	RADISIC M, PARK H, GERECHT S, et al. Biomimetic approach to cardiac tissue engineering[J]. Philos Trans R Soc Lond B Biol Sci, 2007, 362(1484):1357-1368.
14.	李凯, 姜建, 姚晓林, 等.低频脉冲电磁场诱导大鼠骨髓间充质干细胞ACTN2、α-actin和TNNT2表达[J].四川大学学报:医学版, 2012, 43(5):670-674, 710.
15.	况薇, 唐敏, 何学令, 等.周期性拉伸应变对大鼠骨髓间充质干细胞向心肌样细胞分化的影响[J].生物医学工程学杂志, 2014, 31(3):596-600.

1. BARRY F P, MURPHY J M. Mesenchymal stem cells:clinical applications and biological characterization[J]. Int J Biochem Cell Biol, 2004, 36(4):568-584.
2. OHNISHI S, OHGUSHI H, KITAMURA S, et al. Mesenchymal stem cells for the treatment of heart failure[J]. Int J Hematol, 2007, 86(1):17-21.
3. ANTONITSIS P, IOANNIDOU-PAPAGIANNAKI E, KAIDOGLOU A, et al. In vitro cardiomyogenic differentiation of adult human bone marrow mesenchymal stem cells. The role of 5-azacytidine[J]. Interact Cardiovasc Thorac Surg, 2007, 6(5):593-597.
4. RADISIC M, PARK H, SHING H, et al. Functional assembly of engineered myocardium by electrical stimulation of cardiac myocytes cultured on scaffolds[J]. Proc Natl Acad Sci U S A, 2004, 101(52):18129-18134.
5. 严中琴, 杨刚, 崔燎, 等.电刺激促骨髓间充质干细胞向心肌样细胞分化[J].生物医学工程学杂志, 2013, 30(3):556-561.
6. MINGUELL J J, ERICES A, CONGET P. Mesenchymal stem cells[J]. Exp Biol Med (Maywood), 2001, 226(6):507-520.
7. NAGAYA N, FUJⅡT, IWASE T, et al. Intravenous administration of mesenchymal stem cells improves cardiac function in rats with acute myocardial infarction through angiogenesis and myogenesis[J]. Am J Physiol Heart Circ Physiol, 2004, 287(6):H2670-H2676.
8. REBELATTO C K, AGUIAR A M, SENEGAGLIA A C, et al. Expression of cardiac function genes in adult stem cells is increased by treatment with nitric oxide agents[J]. Biochem Biophys Res Commun, 2009, 378(3):456-461.
9. WANG Tingzhong, XU Zhengyun, JIANG Wenhui, et al. Cell-to-cell contact induces mesenchymal stem cell to differentiate into cardiomyocyte and smooth muscle cell[J]. Int J Cardiol, 2006, 109(1):74-81.
10. BURLACU A, ROSCA A M, MANIU H, et al. Promoting effect of 5-azacytidine on the myogenic differentiation of bone marrow stromal cells[J]. Eur J Cell Biol, 2008, 87(3):173-184.
11. MARUTHA RAVINDRAN C R, TICKU M K. Effect of 5-azacytidine on the methylation aspects of NMDA receptor NR2B gene in the cultured cortical neurons of mice[J]. Neurochem Res, 2009, 34(2):342-350.
12. GENOVESE J A, SPADACCIO C, LANGER J, et al. Electrostimulation induces cardiomyocyte predifferentiation of fibroblasts[J]. Biochem Biophys Res Commun, 2008, 370(3):450-455.
13. RADISIC M, PARK H, GERECHT S, et al. Biomimetic approach to cardiac tissue engineering[J]. Philos Trans R Soc Lond B Biol Sci, 2007, 362(1484):1357-1368.
14. 李凯, 姜建, 姚晓林, 等.低频脉冲电磁场诱导大鼠骨髓间充质干细胞ACTN2、α-actin和TNNT2表达[J].四川大学学报:医学版, 2012, 43(5):670-674, 710.
15. 况薇, 唐敏, 何学令, 等.周期性拉伸应变对大鼠骨髓间充质干细胞向心肌样细胞分化的影响[J].生物医学工程学杂志, 2014, 31(3):596-600.

Journal of Biomedical Engineering

Expression of Myocardial Specificity Markers MEF-2C and Cx43 in Rat Bone Marrow-derived Mesenchymal Stem Cells Induced by Electrical Stimulation In Vitro

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content