Cytotechnology

, 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

Abstract

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.

Keywords

Bone marrow Stem cell Cardiomyocyte Differentiation Conditioned media 

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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

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