Cardiogel as an Instructive Microenvironment for in vitro Differentiation of Bone Marrow Derived Mesenchymal Stem Cells into Cardiomyocytes

Background: Stem cell therapy has been developed as an effective treatment method for the heart failure. Also, extracellular matrix has shown the positive effects in stem cell differentiation and myocardial tissue organization. Cardiogel is a native cardiac extracellular matrix (ECM) derived from cardiac fibroblasts. In the present study the role of cardiogel is examined for its cardiomyogenic potential on mouse bone marrow- derived mesenchymal stem cells (BM-MSCs). Method: The BM-MSCs were isolated from six-week-old mice. Cardiac fibroblasts were collected from neonatal heart mice and the cells were seeded on 0.2% gelatin pre-coated plates for up to 21 days. Then, the decellularization was performed via enzymatic digestion. For cardiomyocyte differentiation, the BM-MSCs were plated on matrix-coated plates (Cardiogel, CCP), Matrigel-coated plates (MCP) and gelatin-coated plates (GCP) as a control group at a density of 1 × 10^4 cells per cm² in CM containing 3 μM 5-azacytidine. Results: The results proved that cardiogel is capable to promote the cardiomyogenic differentiation of BM-MSCs. Cardiogel had a great influence on cellular adhesion, gene expression and cardiomyocyte differentiation compared to MCP and GCP. Gene expression analysis showed that cardiac specific markers were expressed in BM-MSCs has grown in CCP that represented the cardiac-specific differentiation. Conclusion: Our results suggest that cardiogel is an effective ECM that can improve the cardiomyogenic differentiation of BM-MSCs, and it can be used as a possible therapeutic approach in cardiac tissue regeneration.