Advertisement

Molecular and Cellular Biochemistry

, Volume 361, Issue 1–2, pp 315–320 | Cite as

In vitro study of the differentiation of bone marrow stromal cells into cardiomyocyte-like cells

  • Karimeh Haghani
  • Salar Bakhtiyari
  • Ali Mohammad Nouri
Article

Abstract

Bone marrow multipotent stromal cells (BMSCs) have the ability to transdifferentiate into various cell types, including: osteoblasts, chondrocytes, adipocytes, neurons, and cardiomyocytes. This study aimed to differentiate the BMSCs into cardiomyocyte. BMSCs were exposed to 5-azacytidine for 24 h. Seven days after the induction of cell differentiation by 5-azacytidine, the cardiomyogenic cells were stained by fushin and binucleated cells were counted and compared with the neonate cardiomyocyte as positive control. In addition, immunofluorescence analysis and western blot were performed using the antibodies against α-actinin, desmin, troponin T, and β-myosin heavy chain. Our results showed that there was no significant difference between the number of binucleated cells within the cardiomyogenic cell group and positive control group; however, a statistically significant difference was observed between both of these groups and undifferentiated cell group (P < 0.005). In addition, after 5-azacytidine treatment, BMSCs had a higher expression of cardiac-specific markers such as desmin, α-actinin, troponin T and β-myosin heavy chain compared with the untreated groups (P < 0.005). We concluded that 5-azacytidine is an effective inducer for the differentiation of BMSCs into cardiomyocytes and could produce a population of binucleated cells, which express α-actinin, desmin, troponin T, and β-myosin heavy chain, four markers of cardiomyocytes.

Keywords

Bone marrow Cardiomyocyte-like cells 5-Azacytidine Desmin α-Actinin 

Notes

Acknowledgments

We thank Dr. Moemeni for his constant encouragement and advice during the course of this work. We also thank Dr. Monavar for his kind helps and comments.

References:

  1. 1.
    Soonpaa MH, Koh GY, Klug MG, Field LJ (1994) Formation of nascent intercalated disks between grafted fetal cardiomyocytes and host myocardium. Science 264:98–101PubMedCrossRefGoogle Scholar
  2. 2.
    Valiunas V, Doronin S, Valiuniene L, Potapova I, Zuckerman J, Walcott B, Robinson RB, Rosen MR, Brink PR, Cohen IS (2004) Human mesenchymal stem cells make cardiac connexins and form functional gap junctions. J Physiol 555:617–626PubMedCrossRefGoogle Scholar
  3. 3.
    Iijima Y, Nagai T, Mizukami M, Matsuura K, Ogura T, Wada H, Toko H, Akazawa H, Takano H, Nakaya H, Komuro I (2003) Beating is necessary for transdifferentiation of skeletal muscle-derived cells into cardiomyocytes. FASEB J 17:1361–1363PubMedGoogle Scholar
  4. 4.
    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 418:41–49PubMedCrossRefGoogle Scholar
  5. 5.
    Friedenstein AJ, Piatetzky S II, Petrakova KV (1966) Osteogenesis in transplants of bone marrow cells. J Embryol Exp Morphol 16:381–390PubMedGoogle Scholar
  6. 6.
    Owen M (1988) Marrow stromal stem cells. J Cell Sci Suppl 10:63–76PubMedGoogle Scholar
  7. 7.
    Pittenger MF, Marshak DR (2001) Mesenchymal stem cells of human adult bone marrow. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  8. 8.
    Bianco P, Cossu G (1999) Uno, nessuno e centomila: searching for the identity of mesodermal progenitors. Exp Cell Res 251:257–263PubMedCrossRefGoogle Scholar
  9. 9.
    Ferrari G, Cusella-De Angelis G, Coletta M, Paolucci E, Stornaiuolo A, Cossu G, Mavilio F (1998) Muscle regeneration by bone marrow-derived myogenic progenitors. Science 279:1528–1530PubMedCrossRefGoogle Scholar
  10. 10.
    Jackson KA, Majka SM, Wang H, Pocius J, Hartley CJ, Majesky MW, Entman ML, Michael LH, Hirschi KK, Goodell MA (2001) Regeneration of ischemic cardiac muscle and vascular endothelium by adult stem cells. J Clin Invest 107:1395–1402PubMedCrossRefGoogle Scholar
  11. 11.
    Orlic D, Kajstura J, Chimenti S, Jakoniuk I, Anderson SM, Li B, Pickel J, McKay R, Nadal-Ginard B, Bodine DM, Leri A, Anversa P (2001) Bone marrow cells regenerate infarcted myocardium. Nature 410:701–705PubMedCrossRefGoogle Scholar
  12. 12.
    Jackson KA, Mi T, Goodell MA (1999) Hematopoietic potential of stem cells isolated from murine skeletal muscle. Proc Natl Acad Sci USA 96:14482–14486PubMedCrossRefGoogle Scholar
  13. 13.
    Zhang FB, Li L, Fang B, Zhu DL, Yang HT, Gao PJ (2005) Passage-restricted differentiation potential of mesenchymal stem cells into cardiomyocyte-like cells. Biochem Biophys Res Commun 336:784–792PubMedCrossRefGoogle Scholar
  14. 14.
    Bakhtiyari S, Meshkani R, Taghikhani M, Larijani B, Adeli K (2010) Protein tyrosine phosphatase-1B (PTP-1B) knockdown improves palmitate-induced insulin resistance in C2C12 skeletal muscle cells. Lipids 45:237–244PubMedCrossRefGoogle Scholar
  15. 15.
    Fukuda K (2001) Development of regenerative cardiomyocytes from mesenchymal stem cells for cardiovascular tissue engineering. Artif Organs 25:187–193PubMedCrossRefGoogle Scholar
  16. 16.
    Chen CJ, Ou YC, Liao SL, Chen WY, Chen SY, Wu CW, Wang CC, Wang WY, Huang YS, Hsu SH (2007) Transplantation of bone marrow stromal cells for peripheral nerve repair. Exp Neurol 204:443–453PubMedCrossRefGoogle Scholar
  17. 17.
    Scorsin M, Marotte F, Sabri A, Le Dref O, Demirag M, Samuel JL, Rappaport L, Menasche P (1996) Can grafted cardiomyocytes colonize peri-infarct myocardial areas? Circulation 94:337–340Google Scholar
  18. 18.
    Watanabe E, Smith DM Jr, Delcarpio JB, Sun J, Smart FW, Van Meter CH Jr, Claycomb WC (1998) Cardiomyocyte transplantation in a porcine myocardial infarction model. Cell Transplant 7:239–246PubMedCrossRefGoogle Scholar
  19. 19.
    Wakitani S, Saito T, Caplan AI (1995) Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve 18:1417–1426PubMedCrossRefGoogle Scholar
  20. 20.
    Makino S, Fukuda K, Miyoshi S, Konishi F, Kodama H, Pan J, Sano M, Takahashi T, Hori S, Abe H, Hata J, Umezawa A, Ogawa S (1999) Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest 103:697–705PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Karimeh Haghani
    • 1
  • Salar Bakhtiyari
    • 1
  • Ali Mohammad Nouri
    • 2
  1. 1.Department of Clinical Biochemistry, Faculty of MedicineIlam University of Medical SciencesIlamIran
  2. 2.Department of Biology, Faculty of Basic SciencesRazi UniversityKermanshahIran

Personalised recommendations