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首页> 外文期刊>Canadian Journal of Physiology and Pharmacology >Timing of induction of cardiomyocyte differentiation for in vitro cultured mesenchymal stem cells: a perspective for emergencies.
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Timing of induction of cardiomyocyte differentiation for in vitro cultured mesenchymal stem cells: a perspective for emergencies.

机译：体外培养的间充质干细胞诱导心肌细胞分化的时机：紧急情况的观点。

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Mesenchymal stem cells (MSCs) have the capacity to differentiate into osteoblasts, chondrocytes, adipocytes, myocytes, and cardiomyocytes. Several established methods are presently available for in vitro isolation of MSCs from bone marrow. However, the duration necessary to culture them can be a major handicap to cell-based therapies needed for such urgent cardiovascular conditions as acute myocardial infarction and acute hindlimb ischemia. The best timing of cardiomyocyte differentiation induction after MCS isolation and expansion is still an unresolved issue. Our goal was to investigate the possibility of obtaining functional cardiomyocytes from rat MSC within a shorter time period. We examined MSCs' colony-forming capacity, CD90 and CD34 immunoreactivity during the 14 days of culturing. Cardiomyocyte differentiation was induced by 5-azacytidine. Immunohistochemic staining, together with intracellular Ca2+ measurement experiments, revealed that MSCs do not differentiate into any specific cell lineage but show the characteristics of MSCs on both the 9th and 14th days of the culture. To check the potential for differentiation into cardiomyocytes, experiments with caffeine application and depolarization with KCl were performed. The cells possessed some of the specific biochemical features of contracting cells, with slightly higher capacities on the 14th day. Cells from 9th and 14th days of the culture that were treated with 5-azacytidine had a higher expression of cardiac-specific markers such as troponin I, alpha-sarcomeric actin, and MEF2D compared with the control groups. This study illustrates that it is possible to get functional cardiomyocytes from in vitro MSC culture in a shorter time period than previously achieved. This reduction in time may provide emergency cases with access to cell-based therapies that may have previously been unavailable.
机译：间充质干细胞（MSC）具有分化为成骨细胞，软骨细胞，脂肪细胞，肌细胞和心肌细胞的能力。目前有几种已建立的方法可用于从骨髓中体外分离MSC。但是，培养它们所需的时间可能是紧急心血管疾病（如急性心肌梗塞和急性后肢缺血）所需的细胞疗法的主要障碍。 MCS分离和扩增后诱导心肌细胞分化的最佳时机仍未解决。我们的目标是研究在较短时间内从大鼠MSC获得功能性心肌细胞的可能性。我们在培养的14天中检查了MSC的集落形成能力，CD90和CD34免疫反应性。 5-氮杂胞苷诱导心肌细胞分化。免疫组织化学染色以及细胞内Ca2 +测量实验表明，MSC在培养的第9天和第14天均未分化为任何特定的细胞谱系，但显示了MSC的特征。为了检查分化为心肌细胞的潜力，进行了咖啡因应用和KCl去极化的实验。这些细胞具有收缩细胞的某些特定生化特征，在第14天的容量稍高。与对照组相比，用5-氮杂胞苷处理的培养的第9天和第14天的细胞具有更高的心脏特异性标志物（如肌钙蛋白I，α-肌氨酸肌动蛋白和MEF2D）表达。这项研究表明，从体外MSC培养中获得功能性心肌细胞的时间比以前要短。时间的减少可以为紧急情况提供以前可能无法获得的基于细胞的疗法。

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