Small molecule 2′-deoxycytidine differentiates human umbilical cord-derived MSCs into cardiac progenitors in vitro and their in vivo xeno-transplantation improves cardiac function
Small molecule 2′-deoxycytidine differentiates human umbilical cord-derived MSCs into cardiac progenitors in vitro and their in vivo xeno-transplantation improves cardiac function
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DOI：
10.1007/s11010-020-03750-6
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发表时间：
2020-05-15
影响因子：
4.3
通讯作者：
Khan, Irfan
中科院分区：
文献类型：
--
作者：
Ali, Syeda Roohina;Ahmad, Waqas;Khan, Irfan
小分子被广泛用于诱导干细胞分化。2 '-脱氧胞苷（2-DC）属于胞苷家族。它刺激心脏特异性基因和蛋白质的表达，并指导间充质干细胞向心肌分化。本研究旨在探讨2-DC诱导的人脐带间充质干细胞（UC-MSCs）向肌源性分化的作用及其在梗死心肌再生中的应用。在细胞毒性分析优化后，用5、10、20和40 μ M 2-DC处理UC-MSC。结扎大鼠冠状动脉左前降支建立心肌梗死（MI）模型。结扎后立即将正常和2-DC处理的UC-MSCs移植到左心室壁中。进行超声心动图测量以评估心脏功能。通过组织学分析检查心肌的组织结构以确定移植细胞的命运。成功地从人脐带组织中分离出了间充质干细胞。在所有浓度下，2-DC处理在UC-MSC中不产生任何显著的细胞毒性作用。经处理的UC-MSC的qPCR分析显示诱导肌源性分化，这在20 μ M浓度下更明显。与正常UC-MSC相比，荧光标记的2-DC处理的UC-MSC显示出在梗死心肌中的显著（**P < 0.01）归巢。与正常UC-MSCs组和MI组相比，移植有2-DC处理的UC-MSCs的心脏显著改善心脏收缩和舒张功能以及泵送能力（*P < 0.001）。与正常UC-MSCs相比，2-DC处理组的纤维化面积和左心室壁厚度显著改善（*P < 0.001）。免疫组织化学染色显示荧光标记的细胞和分化的心肌细胞的斑块的共定位，所述分化的心肌细胞在梗死区中被染色用于心脏蛋白，这意味着经处理的UC-MSC再生心肌细胞。我们首次报道了2-DC诱导UC-MSCs的心脏分化。移植的细胞分化为功能性心肌细胞，并显着改善心脏性能。这些预分化的心脏祖细胞表现出更好的存活、归巢和在梗死区的分布。2-DC处理的细胞不仅改善了心脏功能，而且恢复了组织稳态，这表明在临床设置中心脏组织再生的更好的治疗选择。
Small molecules are widely used to induce stem cell differentiation. 2 '-deoxycytidine (2-DC) belongs to the cytidine family. It stimulates the expression of cardiac-specific genes and proteins, and directs mesenchymal stem cells towards cardiomyogenic differentiation. We aim to investigate the role of 2-DC-treated human umbilical cord mesenchymal stem cells (UC-MSCs) into myogenic lineage and explore their application in regeneration of infarcted myocardium. UC-MSCs were treated with 5, 10, 20, and 40 mu M 2-DC following optimization by cytotoxicity analysis. Rat model of myocardial infarction (MI) was induced by ligating left anterior descending coronary artery. Normal, and 2-DC treated UC-MSCs were transplanted in the left ventricular wall immediately after ligation. Echocardiographic measurements were performed to assess cardiac function. Tissue architecture of the myocardium was examined by histological analysis to determine fate of the transplanted cells. MSCs were successfully isolated from human umbilical cord tissue. 2-DC treatment did not produce any significant cytotoxic effect in UC-MSCs at all concentrations. qPCR analysis of treated UC-MSCs showed induction of myogenic differentiation, which is more pronounced at 20 mu M concentration. Fluorescently labeled 2-DC-treated UC-MSCs showed significant (**P < 0.01) homing in the infarcted myocardium as compared to normal UC-MSCs. Hearts transplanted with 2-DC-treated UC-MSCs significantly (***P < 0.001) improved the cardiac systolic and diastolic functions and pumping ability as compared to normal UC-MSCs and MI groups. Fibrotic area and left ventricular wall thickness were significantly improved (***P < 0.001) in 2-DC-treated group as compared to normal UC-MSCs. Immunohistochemical staining showed co-localization of fluorescently labeled cells and patches of differentiated myocytes which were stained for cardiac proteins in the infarct zone implying that the treated UC-MSCs regenerated cardiomyocytes. We report for the first time that 2-DC induces cardiac differentiation in UC-MSCs. Transplanted cells differentiated into functional cardiomyocytes and significantly improved cardiac performance. These pre-differentiated cardiac progenitors showed better survival, homing, and distribution in the infarcted zone. 2-DC treated cells not only improved cardiac function, but also restored tissue homeostasis, suggesting a better therapeutic option for the regeneration of cardiac tissue in the clinical setup.