, Volume 64, Issue 5, pp 563–575 | Cite as

Ischemic cardiac tissue conditioned media induced differentiation of human mesenchymal stem cells into early stage cardiomyocytes

  • Balasundari Ramesh
  • Dillip Kumar Bishi
  • Suneel Rallapalli
  • Sarasabarathi Arumugam
  • Kotturathu Mammen Cherian
  • Soma Guhathakurta
Original Research


Mesenchymal stem cells (MSCs) are multipotent, can be easily expanded in culture and hence are an attractive therapeutic tool for cardiac repair. MSCs have tremendous potential to transdifferentiate to cardiac lineage both in vitro and in vivo. The present study examined the differentiation capacity of conditioned media derived from ischemic cardiac tissue on human MSCs. Human Bone marrow-derived MSCs after due characterization by immunocytochemistry and flow cytometry for MSC specific markers were induced by culture media derived from ischemic (n = 13) and non-ischemic (n = 18) human cardiac tissue. Parallel cultures were treated with 5-azacytidine (5-azaC), a potent cardiomyogen. MSCs induced with ischemic conditioned media formed myotube like structures, expressed sarcomeric Troponin I, alpha myosin heavy chain proteins and were positive for cardiac specific markers (Nkx2.5, human atrial natriuretic peptide, myosin light chain-2a, GATA-4) as was observed in 5-azaC treated cells. However, uninduced MSCs as well as those induced with non-ischemic cardiac conditioned media still maintained the fibroblast morphology even after 3 weeks post-induction. Transmission electron microscopic studies of cardiomyocyte-like cells derived from MSCs revealed presence of sarcomeric bands but failed to show gap junctions and intercalated discs as of adult cardiomyocytes. These findings demonstrate that ischemic cardiac conditioned media induces morphological and molecular changes in MSCs with cardiac features, but at a primitive stage. Proteomics analysis of the ischemic conditioned media revealed differential expression of three relevant proteins (C-type lectin superfamily member 13, Testis-specific chromodomain protein Y2 and ADP/ATP translocase 1), whose exact role in cardiac regeneration needs further analysis.


Bone marrow Stem cell Cardiomyocyte Differentiation Conditioned media 


  1. Antonitsis P, Ioannidou-Papagiannaki E, Kaidoglou A, Papakonstantinou C (2007) In vitro cardiomyogenic differentiation of adult human bone marrow mesenchymal stem cells. The role of 5-azacytidine. Interact Cardiovasc Thorac Surg 6:593–597CrossRefGoogle Scholar
  2. Beltrami AP, Urbanek K, Kajustura J, Yan SM, Finato N, Bussani R, Nadal-Ginard B, Silvestri F, Leri A, Beltrami A, Anversa P (2001) Evidence that human cardiac myocytes divide after myocardial infarction. N Engl J Med 344:1750–1757CrossRefGoogle Scholar
  3. Chang SA, Lee EJ, Kang HJ, Zhang SY, Kim JH, Li L, Youn SW, Lee CS, Kim KH, Won JY, Sohn JW, Park KW, Cho HJ, Yang SE, Oh WI, Yang YS, Ho WK, Park YB, Kim HS (2008) Impact of myocardial infarct proteins and oscillating pressure on the differentiation of mesenchymal stem cells: effect of acute myocardial infarction on stem cell differentiation. Stem Cells 26:1901–1912Google Scholar
  4. Fraser JK, Schreiber RE, Zuk PA, Hedrick MH (2004) Adult stem cell therapy for the heart. Int J Biochem Cell Biol 36:658–666CrossRefGoogle Scholar
  5. Fukuda K, Yuasa S (2003) Stem cells as a source of regenerative cardiomyocytes use of adult marrow mesenchymal stem cells for regeneration of cardiomyocytes. Bone Marrow Transplant 32:S25–S27CrossRefGoogle Scholar
  6. Hassink RJ, Brutel de la Rivière A, Mummery CL, Doevendans PA (2003) Transplantation of cells for cardiac repair. J Am Coll Cardiol 41:711–717CrossRefGoogle Scholar
  7. Heng BC, Haider HK, Sim EK, Cao T, Ng SC (2004) Strategies for directing the differentiation of stem cells into the cardiomyogenic lineage in vitro. Cardiovasc Res 62:34–42CrossRefGoogle Scholar
  8. Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, Reyes M, Lenvik T, Lund T, Blackstad M, Du J, Aldrich S, Lisberg A, Low WC, Largaespada DA, Verfaillie CM (2002) Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418:41–49Google Scholar
  9. Juttermann R, Li E, Jaenisch R (1994) Toxicity of 5-axa-2-deoxycytidine to mammalian cells is mediated primarily by co-valent trapping of DNA methyl transferase rather than DNA demethylation. Proc Natl Acad Sci USA 19:11797–11801CrossRefGoogle Scholar
  10. Kamihata H, Matsubara H, Nishiue T, Fujiyama S, Tsutsumi Y, Ozono R, Masaki H, Mori Y, Iba O, Tateishi E, Kosaki A, Shintani S, Murohara T, Imaizumi T, Iwasaka T (2001) Implantation of bone marrow mononuclear cells into ischemic myocardium enhances collateral perfusion and regional function via side supply of angioblasts, angiogenic ligands, and cytokines. Circulation 104:1046–1052Google Scholar
  11. Kato M, Khan S, d’Aniello E, McDonald KJ, Hart DN (2007) The novel endocytic and phagocytic D-type lectin receptor DCL-1/CD302 on macrophages is colacalized with F-Actin, sugessing a role in cell adhesion and migration. J Immunol 179:6052–6063Google Scholar
  12. Lee SH, Wolf PL, Escudero R, Deutsch R, Jamieson SW, Thistlethwaite PA (2000) Early expression of angiogenesis factors in acute myocardial ischemia and infarction. N Engl J Med 342:626–633CrossRefGoogle Scholar
  13. Lunardi J, Hurko O, Engel WK, Attardi G (1992) The multiple ADP/ATP translocase genes are differentially expressed during human muscle development. J Biol Chem 267:15267–15270Google Scholar
  14. Mariani MA, Alfonso AD, Croccia M, Limbruno U, Stefano RD, Grandjean JG (2004) Stem cell transplantation for ischemic myocardium. Ital Heart J 5:340–342Google Scholar
  15. Ohnishi S, Nagaya N (2007) Prepare cells to repair heart: mesenchymal stem cells for treatment of heart failure. Am J Nephrol 27:301–307CrossRefGoogle Scholar
  16. Paquin J, Danalache BA, Jankowski M, Mc Cann SM, Gutkowska J (2002) Oxytocin induces differentiation of P19 embryonic stem cells to cardiomyocytes. Proc Natl Acad Sci USA 99:9550–9555CrossRefGoogle Scholar
  17. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147Google Scholar
  18. Rose RA, Jiang H, Wang X, Helke S, Tsoporis JN, Gong N, Keating SC, Parker TG, Backx PH, Keating A (2008) Bone marrow-derived mesenchymal stromal cells express cardiac-specific markers retain the stromal phenotype and do not become functional cardiomyocytes invitro. Stem Cells 26:2884–2892Google Scholar
  19. Sharpe N (2004) Cardiac remodeling in coronary artery disease. Am J Cardiol 93:17B–20BCrossRefGoogle Scholar
  20. Toma C, Pittenger MF, Cahill KS, Byrne BJ, Kessler PD (2002) Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation 105:93–98CrossRefGoogle Scholar
  21. Tyndall A, Walker UA, Cope A, Dazzi F, De Bari C, Fibbe W, Guiducci S, Jones S, Jorgensen C, Le Blanc K, Luyten F, McGonagle D, Martin I, Bocelli-Tyndall C, Pennesi G, Pistoia V, Pitzalis C, Uccelli A, Wulffraat N, Feldmann M (2007) Immunomodulatory properties of mesenchymal stem cells: a review based on an interdisciplinary meeting held at the Kennedy institute of rheumatology division London UK. 31 October 2005. Arthritis Res Ther 9:301Google Scholar
  22. Ventura C, Cantoni S, Bianchi F (2007) Hyaluronan mixed esters of butyric and retinoic acid drive cardiac and endothelial fate in term placenta human mesenchymal stem cells and enhance cardiac repair in infracted rat hearts. J Biol Chem 282:14243–14252CrossRefGoogle Scholar
  23. Weissberg PL, Qasim A (2005) Stem cell therapy for myocardial repair. Heart 91:696–702CrossRefGoogle Scholar
  24. Wu H, Min J, Antoshenko T, Plotnikov AN (2009) Crystal structures of human CDY proteins reveal a crotonase-like fold proteins. Proteins 76:1054–1061CrossRefGoogle Scholar
  25. Zhang SX, Garcia-Gras E, Wycuff DR, Marriot SJ, Kadeer N, Yu W, Olson EN, Garry DJ, Parmacek MS, Schwartz RJ (2005) Identification of direct serum-response factor gene targets during Me2SO-induced P19 cardiac cell differentiation. J Biol Chem 280:19115–19126Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Balasundari Ramesh
    • 1
  • Dillip Kumar Bishi
    • 1
  • Suneel Rallapalli
    • 1
  • Sarasabarathi Arumugam
    • 2
  • Kotturathu Mammen Cherian
    • 1
  • Soma Guhathakurta
    • 1
  1. 1.International Centre for Cardio Thoracic and Vascular DiseasesFrontier Lifeline and Dr. K.M.Cherian Heart FoundationChennaiIndia
  2. 2.MIOT HospitalChennaiIndia

Personalised recommendations