Farag Ahmed, Koung Ngeun Sai, Kaneda Masahiro, Aboubakr Mohamed, Tanaka Ryou
Veterinary Teaching Hospital, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
Department of Surgery, Anesthesiology, and Radiology, Faculty of Veterinary Medicine, Zagazig University, Zagazig 44519, Egypt.
Biomedicines. 2024 Aug 22;12(8):1923. 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.
间充质干细胞（MSCs）具有多能性、自我更新能力和免疫调节特性，使其在再生医学中，特别是在心血管治疗方面具有广阔前景。然而，优化MSCs的来源和心脏分化的诱导方法具有挑战性。本研究比较了单独使用5-氮杂胞苷（5-Aza）或与低剂量成纤维细胞生长因子（FGF）和胰岛素样生长因子（IGF）联合使用时骨髓（BM）-MSCs和脂肪来源（AD）-MSCs的心肌生成潜力。BM-MSCs和AD-MSCs采用两种方案进行分化：单独使用10 μmol 5-Aza以及10 μmol 5-Aza与1 ng/mL FGF和10 ng/mL IGF联合使用。进行了形态学、转录和翻译分析以及细胞活力评估。两种类型的MSCs都表现出相似的形态变化；然而，AD-MSCs比BM-MSCs更快达到70-80%的汇合度。表面标志物分析证实了CD29和CD90呈阳性，CD45呈阴性。分化方案导致细胞变平并形成肌管，AD-MSCs的分化更早。联合方案降低了BM-MSCs的细胞死亡率，并增强了心脏标志物（……）的表达，特别是在BM-MSCs中。免疫荧光证实了所有处理组中均有心脏特异性蛋白表达。两种类型的MSCs都表现出指示心肌分化的心脏特异性标志物的表达，联合处理对BM-MSCs显示出更高的效率。
