Molecular and Cellular Biochemistry

, Volume 339, Issue 1–2, pp 89–98 | Cite as

Co-culture with cardiomyocytes enhanced the myogenic conversion of mesenchymal stromal cells in a dose-dependent manner

  • Xiao-qing He
  • Min-sheng Chen
  • Shu-Hong Li
  • Shi-ming Liu
  • Yun Zhong
  • Heather Y. McDonald Kinkaid
  • Wei-Yang Lu
  • Richard D. Weisel
  • Ren-Ke Li


To increase the accessibility of myogenic cells for cell therapy in the infarcted heart, we identified conditions to improve the reproducible conversion of bone marrow mesenchymal stromal cells (BMSCs) into myogenic cells. Such cells may permit functional regeneration following a myocardial infarction. BMSCs derived from green fluorescent protein (GFP) transgenic rats were co-cultured with neonatal rat cardiomyocytes (1:1, 1:10, 1:20, and 1:40 ratios) for 7 days. Some BMSCs contracted synchronously with the neonatal cardiomyocytes, and exhibited action potentials that were confirmed with current clamp recordings. The myogenic phenotype of the BMSCs was confirmed by immunohistochemical staining and flow cytometry (antibodies against cardiac specific α-sarcomeric actinin, Troponin I, MEF-2C). An increase in the number of BMSCs expressing cardiac markers correlated with increasing numbers of neonatal cardiomyocytes in the culture. When BMSCs were co-cultured with DiI-labeled neonatal cardiomyocytes, a small percentage of GFP/DiI/Troponin I triple-positive cells were observed after 7 days. This type of myogenic conversion increased nearly twofold when BMSCs were co-cultured with apoptotic (TNF-α-treated) cardiomyocytes. BMSCs co-cultured with cardiomyocytes acquired a functional myogenic phenotype in a dose-dependent manner. Myogenic conversion increased when the BMSCs were cultured with apoptotic cells.


Bone marrow stromal cells Cardiomyocytes Cell fusion Co-culture Differentiation Myogenic 



This study was supported by the Heart and Stroke Foundation of Ontario (T6604 to RL), and the Guangdong Provincial International Science and Technology Cooperation Foundation (2006B50107005 to MC). RL is a Career Investigator of the Heart and Stroke Foundation of Canada, and holds a Canada Research Chair in Cardiac Regeneration. MC is a Peer Reviewer of the National Natural Science Foundation of China and a standing director of the Guangdong Medical Doctors Association.


  1. 1.
    American Heart Association (2001) Heart and stroke statistical update. Dallas, TX, American Heart AssociationGoogle Scholar
  2. 2.
    Dowell JD, Rubart M, Pasumarthi KB, Soonpaa MH, Field LJ (2003) Myocyte and myogenic stem cell transplantation in the heart. Cardiovasc Res 58:336–350CrossRefPubMedGoogle Scholar
  3. 3.
    Itescu S, Schuster MD, Kocher AA (2003) New directions in strategies using cell therapy for heart disease. J Mol Med 81:288–296PubMedGoogle Scholar
  4. 4.
    Dawn B, Bolli R (2005) Bone marrow cells for cardiac regeneration: the quest for the protagonist continues. Cardiovasc Res 65:293–295CrossRefPubMedGoogle Scholar
  5. 5.
    Zwaginga JJ, Doevendans P (2003) Stem cell-derived angiogenic/vasculogenic cells: possible therapies for tissue repair and tissue engineering. Clin Exp Pharmacol Physiol 30:900–908CrossRefPubMedGoogle Scholar
  6. 6.
    Pittenger MF, Martin BJ (2004) Mesenchymal stem cells and their potential as cardiac therapeutics. Circ Res 95:9–20CrossRefPubMedGoogle Scholar
  7. 7.
    Makino S, Fukuda K, Miyoshi S et al (1999) Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest 103:697–705CrossRefPubMedGoogle Scholar
  8. 8.
    Rosenblatt-Velin N, Lepore MG, Cartoni C, Beermann F, Pedrazzini T (2005) FGF-2 controls the differentiation of resident cardiac precursors into functional cardiomyocytes. J Clin Invest 115:1724–1733CrossRefPubMedGoogle Scholar
  9. 9.
    Fukuda K, Fujita J (2005) Mesenchymal, but not hematopoietic, stem cells can be mobilized and differentiate into cardiomyocytes after myocardial infarction in mice. Kidney Int 68:1940–1943CrossRefPubMedGoogle Scholar
  10. 10.
    Kawada H, Fujita J, Kinjo K et al (2004) Nonhematopoietic mesenchymal stem cells can be mobilized and differentiate into cardiomyocytes after myocardial infarction. Blood 104:3581–3587CrossRefPubMedGoogle Scholar
  11. 11.
    Fukuhara S, Tomita S, Yamashiro S et al (2003) Direct cell-cell interaction of cardiomyocytes is key for bone marrow stromal cells to go into cardiac lineage in vitro. J Thorac Cardiovasc Surg 125:1470–1480CrossRefPubMedGoogle Scholar
  12. 12.
    Rangappa S, Entwistle JW, Wechsler AS, Kresh JY (2003) Cardiomyocyte-mediated contact programs human mesenchymal stem cells to express cardiogenic phenotype. J Thorac Cardiovasc Surg 126:124–132CrossRefPubMedGoogle Scholar
  13. 13.
    Sakai T, Li RK, Weisel RD et al (1999) Fetal cell transplantation: a comparison of three cell types. J Thorac Cardiovasc Surg 118:715–724CrossRefPubMedGoogle Scholar
  14. 14.
    Waspe LE, Ordahl CP, Simpson PC (1990) The cardiac beta-myosin heavy chain isogene is induced selectively in alpha 1-adrenergic receptor-stimulated hypertrophy of cultured rat heart myocytes. J Clin Invest 85:1206–1214CrossRefPubMedGoogle Scholar
  15. 15.
    Lu WY, Xiong ZG, Lei S et al (1999) G-protein-coupled receptors act via protein kinase C and Src to regulate NMDA receptors. Nat Neurosci 2:331–338CrossRefPubMedGoogle Scholar
  16. 16.
    Alvarez-Dolado M, Pardal R, Garcia-Verdugo JM et al (2003) Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes. Nature 425:968–973CrossRefPubMedGoogle Scholar
  17. 17.
    Terada N, Hamazaki T, Oka M et al (2002) Bone marrow cells adopt the phenotype of other cells by spontaneous cell fusion. Nature 416:542–545CrossRefPubMedGoogle Scholar
  18. 18.
    Fukuda K (2003) Use of adult marrow mesenchymal stem cells for regeneration of cardiomyocytes. Bone Marrow Transplant 32(Suppl 1):S25–S27CrossRefPubMedGoogle Scholar
  19. 19.
    Lagostena L, Avitabile D, De FE et al (2005) Electrophysiological properties of mouse bone marrow c-kit + cells co-cultured onto neonatal cardiac myocytes. Cardiovasc Res 66:482–492CrossRefPubMedGoogle Scholar
  20. 20.
    Murry CE, Soonpaa MH, Reinecke H et al (2004) Haematopoietic stem cells do not transdifferentiate into cardiac myocytes in myocardial infarcts. Nature 428:664–668CrossRefPubMedGoogle Scholar
  21. 21.
    Schafer R, Northoff H (2008) Cardioprotection and cardiac regeneration by mesenchymal stem cells. Panminerva Med 50:31–39PubMedGoogle Scholar
  22. 22.
    Chedrawy EG, Wang JS, Nguyen DM, Shum-Tim D, Chiu RC (2002) Incorporation and integration of implanted myogenic and stem cells into native myocardial fibers: anatomic basis for functional improvements. J Thorac Cardiovasc Surg 124:584–590CrossRefPubMedGoogle Scholar
  23. 23.
    Schulze M, Belema-Bedada F, Technau A, Braun T (2005) Mesenchymal stem cells are recruited to striated muscle by NFAT/IL-4-mediated cell fusion. Genes Dev 19:1787–1798CrossRefPubMedGoogle Scholar
  24. 24.
    Jiang Y, Jahagirdar BN, Reinhardt RL et al (2002) Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418:41–49CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Xiao-qing He
    • 1
    • 2
  • Min-sheng Chen
    • 3
  • Shu-Hong Li
    • 1
  • Shi-ming Liu
    • 4
  • Yun Zhong
    • 1
    • 4
  • Heather Y. McDonald Kinkaid
    • 1
  • Wei-Yang Lu
    • 5
  • Richard D. Weisel
    • 1
  • Ren-Ke Li
    • 1
    • 6
  1. 1.Division of Cardiovascular Surgery and Department of SurgeryToronto General Research Institute and University of TorontoTorontoCanada
  2. 2.Department of CardiologyThe Third Affiliated Hospital of Guangzhou Medical CollegeGuangzhouChina
  3. 3.Guangzhou Medical CollegeGuangzhouChina
  4. 4.Department of CardiologyThe Second Affiliated Hospital of Guangzhou Medical CollegeGuangzhouChina
  5. 5.Department of Physiology and PharmacologyThe University of Western OntarioLondonCanada
  6. 6.MaRS Centre, Toronto Medical Discovery TowerTorontoCanada

Personalised recommendations