Optimizing Cardiomyocyte Differentiation: Comparative Analysis of Bone Marrow and Adipose-Derived Mesenchymal Stem Cells in Rats Using 5-Azacytidine and Low-Dose FGF and IGF Treatment

• Published in: Biomedicines 2024-08, Vol.12 (8), p.1923
• Main Authors: Farag, Ahmed, Koung Ngeun, Sai, Kaneda, Masahiro, Aboubakr, Mohamed, Tanaka, Ryou
• Format: Article
• Language: English
• Subjects: 5-Azacytidine; Amino acids; Antibodies; Azacytidine; Body fat; Bone marrow; Cardiomyocytes; cardiomyogenic differentiation; CD29 antigen; CD45 antigen; Cell differentiation; Cell self-renewal; Cell viability; Comparative analysis; Dosage and administration; Fibroblast growth factors; Fibroblasts; Flow cytometry; growth factors; Health aspects; Heart; Heart attacks; Heart cells; Immunofluorescence; Immunomodulation; Insulin-like growth factor 1; Insulin-like growth factors; Medical research; Medicine, Experimental; Mesenchymal stem cells; Methods; Morphology; Myotubes; Penicillin; Physiological aspects; Regenerative medicine; Scientific equipment and supplies industry; Stem cells; Surface markers; Tissue engineering; Troponin I
• ISSN: 2227-9059; 2227-9059
• DOI: 10.3390/biomedicines12081923

Mesenchymal stem cells (MSCs) exhibit multipotency, self-renewal, and immune-modulatory properties, making them promising in regenerative medicine, particularly in cardiovascular treatments. However, optimizing the MSC source and induction method of cardiac differentiation is challenging. This study compares the cardiomyogenic potential of bone marrow (BM)-MSCs and adipose-derived (AD)-MSCs using 5-Azacytidine (5-Aza) alone or combined with low doses of Fibroblast Growth Factor (FGF) and Insulin-like Growth Factor (IGF). BM-MSCs and AD-MSCs were differentiated using two protocols: 10 μmol 5-Aza alone and 10 μmol 5-Aza with 1 ng/mL FGF and 10 ng/mL IGF. Morphological, transcriptional, and translational analyses, along with cell viability assessments, were performed. Both the MSC types exhibited similar morphological changes; however, AD-MSCs achieved 70-80% confluence faster than BM-MSCs. Surface marker profiling confirmed CD29 and CD90 positivity and CD45 negativity. The differentiation protocols led to cell flattening and myotube formation, with earlier differentiation in AD-MSCs. The combined protocol reduced cell mortality in BM-MSCs and enhanced the expression of cardiac markers ( , , ), particularly in BM-MSCs. Immunofluorescence confirmed cardiac-specific protein expression in all the treated groups. Both MSC types exhibited the expression of cardiac-specific markers indicative of cardiomyogenic differentiation, with the combined treatment showing superior efficiency for BM-MSCs.