DYNAMIC CHANGES OF GENE EXPRESSION PROFILES DURING CARDIOMYOGENESIS OF HUMANMARROW MESENCHYMAL STEM CELLS/_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

YANG Lin ¹ , SHEN Ti ² , CHEN Lianfeng ² , CAO Xinxin ² , LAI Jinzhi. ³

1Department of Oncology, Tongji Hospital of Tongji Medical College, Huazhong University of Science &;
Technology, Wuhan Hubei, 430030,P.R.China;;
2Department of Hematology, 3Department of Cardiovascular Medicine, Peking Union Medical College Hospital, PekingUnion Medical College &;
Chinese Academy of Medical Sciences.;

Corresponding author：

Keywords：

Human bone marrow mesenchymal stem cells ,Gene expression profile Induction, Cardiomyogenic cell

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To analyze the changes of gene expression profiles during the process that human bone
marrow mesenchymal stem cells (hBMSCs) are induced to differentiate into cardiomyogenic cells with 5-azacytidine (5-
aza). Methods hBMSCs were isolated from marrow of obsolete ribs and induced with 5-aza. Then immunocytochemical
staining was used to detect the expressions of α-actin, cardiac troponin T (cTnT), and connexin 43, and the percentage of
cTnT positive cells was tested with flow cytometry. In the process of differentiation, variation of gene expression was screened
with Genechi ps Operating System of human gene expression profiles. And the differentially expressed genes were functionally
analyzed and hierarchical clustered. Results When BMSCs were induced in vitro with 5-aza, part of the cells turned
into myogenic cells morphologically. Before induction, immunocytochemical staining for α-actin and cTnT showed sl ight
positive and for connexin 43 showed negative. While after 3 weeks of induction, immunocytochemical staining for α-actin,
cTnT, and connexin 43 showed all positive. With flow cytometry, the percentage of cTnT positive cells was 7.43% ± 0.02%
before induction, but it was 49.64% ± 0.05% after induction. During differentiation, 1 814 differentially expressed genes
were reported by gene chi ps. Of them, 647 genes were divided into 5 groups with hierarchical clustering. They had various
biological functions, involving signal transduction, cell metabol ism, prol iferation, differentiation, development, and
topogenesis. Conclusion hBMSCs can differentiate into cardiomyogenic cells with the induction of 5-aza in vitro. Multi ple
genes related with signal transduction, transcri ption, and growth factors are involved during this process.

Citation： YANG Lin,SHEN Ti,CHEN Lianfeng,CAO Xinxin,LAI Jinzhi.. DYNAMIC CHANGES OF GENE EXPRESSION PROFILES DURING CARDIOMYOGENESIS OF HUMANMARROW MESENCHYMAL STEM CELLS/. Chinese Journal of Reparative and Reconstructive Surgery, 2012, 26(5): 607-611. doi: Copy

1.	Passier R, van Laake LW, Mummery CL. Stem-cell-based therapy and lessons from the heart. Nature, 2008, 453(7193): 322-329.
2.	Wollert KC, Meyer GP, Lotz J, et al. Intracoronary autologous bone-marrow cell transfer after myocardial infarction: the BOOST randomised controlled clinical trial. Lancet, 2004, 364(9429): 141-148.
3.	Britten MB, Abolmaali ND, Assmus B, et al. Infarct remodeling after intracoronary progenitor cell treatment in patients with acute myocardial infarction (TOPCARE-AMI): mechanistic insights from serial contrast-enhanced magnetic resonance imaging. Circulation, 2003, 108(18): 2212-2218.
4.	Dimmeler S, Zeiher AM, Schneider MD. Unchain my heart: the scientific foundations of cardiac repair. J Clin Invest, 2005, 115(3): 572-583.
5.	Ling SK, Wang R, Dai ZQ, et al. Pretreatment of rat bone marrow mesenchymal stem cells with a combination of hypergravity and 5-azacytidine enhances therapeutic efficacy for myocardial infarction. Biotechnol Prog, 2011, 27(2): 473-482.
6.	Xu W, Zhang X, Qian H, et al. Mesenchymal stem cells from adult human bone marrow differentiates into a cardiomyocyte phenotype in vitro. Exp Biol Medi (Maywood), 2004, 229(7): 623-631.
7.	Makino S, Fukuda K, Miyoshi S, et al. Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest, 1999, 103(5): 697-705.
8.	Cohen ED, Tian Y, Morrisey EE. Wnt signaling: an essential regulator of cardiovascular differentiation, morphogenesis and progenitor self-renewal. Development, 2008, 135(5): 789-798.
9.	Flaherty MP, Abdel-Latif A, Li Q, et al. Noncanonical Wnt11 signaling is sufficient to induce cardiomyogenic differentiation in unfractionated bone marrow mononuclear cells. Circulation, 2008, 117(17): 2241-2252.
10.	Qyang Y, Martin-Puig S, Chiravuri M, et al. The renewal and differentiation of isl1+ cardiovascular progenitors are controlled by a Wnt/beta-catenin pathway. Cell Stem Cell, 2007, 1(2): 165-179.
11.	Zhu W, Shiojima I, Ito Y, et al. IGFBP-4 is an inhibitor of canonical Wnt signalling required for cardiogenesis. Nature, 2008, 454(7202): 345-349.
12.	Schneider VA, Mercola M. Wnt antagonism initiates cardiogenesis in Xenopus laevis. Genes Dev, 2001, 15(3): 304-315.
13.	Tzahor E, Lassar AB. Wnt signals from the neural tube block ectopic cardiogenesis. Genes Dev, 2001, 15(3): 255-260.
14.	Li Y, Hiroi Y, Ngoy S, et al. Notch1 in bone marrow-derived cells mediates cardiac repair after myocardial infarction. Circulation, 2011, 123(8): 866-876.
15.	Crispino JD, Lodish MB, Thurberg BL, et al. Proper coronary vascular development and heart morphogenesis depend on interaction of GATA-4 with FOG cofactors. Genes Dev, 2001, 15(7): 839-844.
16.	Zhao R, Watt AJ, Battle MA, et al. Loss of both GATA4 and GATA6 blocks cardiac myocyte differentiation and results in acardia in mice. Dev Biol, 2008, 317(2): 614-619.
17.	Sachinidis A, Kolossov E, Fleischmann BK, et al. Generation of cardiomyocytes from embryonic stem cells experimental studies. Herz, 2002, 27(7): 589-597.
18.	Yang HS, Bhang SH, Kim IK, et al. In situ cardiomyogenic differentiation of implanted bone marrow mononuclear cells by local delivery of transforming growth factor-β1. Cell Transplant, 2011. [Epub ahead of print].
19.	Wan CR, Chung S, Kamm RD. Differentiation of embryonic stem cells into cardiomyocytes in a compliant microfluidic system. Ann Biomed Eng, 2011, 39(6): 1840-1847.
20.	Afouda BA, Hoppler S. Different requirements for GATA factors in cardiogenesis are mediated by non-canonical Wnt signaling. Dev Dyn, 2011, 240(3): 649-662.

1. Passier R, van Laake LW, Mummery CL. Stem-cell-based therapy and lessons from the heart. Nature, 2008, 453(7193): 322-329.
2. Wollert KC, Meyer GP, Lotz J, et al. Intracoronary autologous bone-marrow cell transfer after myocardial infarction: the BOOST randomised controlled clinical trial. Lancet, 2004, 364(9429): 141-148.
3. Britten MB, Abolmaali ND, Assmus B, et al. Infarct remodeling after intracoronary progenitor cell treatment in patients with acute myocardial infarction (TOPCARE-AMI): mechanistic insights from serial contrast-enhanced magnetic resonance imaging. Circulation, 2003, 108(18): 2212-2218.
4. Dimmeler S, Zeiher AM, Schneider MD. Unchain my heart: the scientific foundations of cardiac repair. J Clin Invest, 2005, 115(3): 572-583.
5. Ling SK, Wang R, Dai ZQ, et al. Pretreatment of rat bone marrow mesenchymal stem cells with a combination of hypergravity and 5-azacytidine enhances therapeutic efficacy for myocardial infarction. Biotechnol Prog, 2011, 27(2): 473-482.
6. Xu W, Zhang X, Qian H, et al. Mesenchymal stem cells from adult human bone marrow differentiates into a cardiomyocyte phenotype in vitro. Exp Biol Medi (Maywood), 2004, 229(7): 623-631.
7. Makino S, Fukuda K, Miyoshi S, et al. Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest, 1999, 103(5): 697-705.
8. Cohen ED, Tian Y, Morrisey EE. Wnt signaling: an essential regulator of cardiovascular differentiation, morphogenesis and progenitor self-renewal. Development, 2008, 135(5): 789-798.
9. Flaherty MP, Abdel-Latif A, Li Q, et al. Noncanonical Wnt11 signaling is sufficient to induce cardiomyogenic differentiation in unfractionated bone marrow mononuclear cells. Circulation, 2008, 117(17): 2241-2252.
10. Qyang Y, Martin-Puig S, Chiravuri M, et al. The renewal and differentiation of isl1+ cardiovascular progenitors are controlled by a Wnt/beta-catenin pathway. Cell Stem Cell, 2007, 1(2): 165-179.
11. Zhu W, Shiojima I, Ito Y, et al. IGFBP-4 is an inhibitor of canonical Wnt signalling required for cardiogenesis. Nature, 2008, 454(7202): 345-349.
12. Schneider VA, Mercola M. Wnt antagonism initiates cardiogenesis in Xenopus laevis. Genes Dev, 2001, 15(3): 304-315.
13. Tzahor E, Lassar AB. Wnt signals from the neural tube block ectopic cardiogenesis. Genes Dev, 2001, 15(3): 255-260.
14. Li Y, Hiroi Y, Ngoy S, et al. Notch1 in bone marrow-derived cells mediates cardiac repair after myocardial infarction. Circulation, 2011, 123(8): 866-876.
15. Crispino JD, Lodish MB, Thurberg BL, et al. Proper coronary vascular development and heart morphogenesis depend on interaction of GATA-4 with FOG cofactors. Genes Dev, 2001, 15(7): 839-844.
16. Zhao R, Watt AJ, Battle MA, et al. Loss of both GATA4 and GATA6 blocks cardiac myocyte differentiation and results in acardia in mice. Dev Biol, 2008, 317(2): 614-619.
17. Sachinidis A, Kolossov E, Fleischmann BK, et al. Generation of cardiomyocytes from embryonic stem cells experimental studies. Herz, 2002, 27(7): 589-597.
18. Yang HS, Bhang SH, Kim IK, et al. In situ cardiomyogenic differentiation of implanted bone marrow mononuclear cells by local delivery of transforming growth factor-β1. Cell Transplant, 2011. [Epub ahead of print].
19. Wan CR, Chung S, Kamm RD. Differentiation of embryonic stem cells into cardiomyocytes in a compliant microfluidic system. Ann Biomed Eng, 2011, 39(6): 1840-1847.
20. Afouda BA, Hoppler S. Different requirements for GATA factors in cardiogenesis are mediated by non-canonical Wnt signaling. Dev Dyn, 2011, 240(3): 649-662.

Chinese Journal of Reparative and Reconstructive Surgery

DYNAMIC CHANGES OF GENE EXPRESSION PROFILES DURING CARDIOMYOGENESIS OF HUMANMARROW MESENCHYMAL STEM CELLS/

Abstract Full text Figures/Tables Video References Cited by

Format

Content