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首页> 外文期刊>Regenerative Therapy >Extracellular matrix from decellularized mesenchymal stem cells improves cardiac gene expressions and oxidative resistance in cardiac C-kit cells
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Extracellular matrix from decellularized mesenchymal stem cells improves cardiac gene expressions and oxidative resistance in cardiac C-kit cells

机译：去细胞的间充质干细胞的细胞外基质改善心脏C-kit细胞中的心脏基因表达和抗氧化性

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Objective Myocardial infarction remains the number one killer disease worldwide. Cellular therapy using cardiac c-kit cells (CCs) are capable of regenerating injured heart. Previous studies showed mesenchymal stem cell-derived (MSC) extracellular matrices can provide structural support and are capable of regulating stem cell functions and differentiation. This study aimed to evaluate the effects of human MSC-derived matrices for CC growth and differentiation. Methods Human Wharton's Jelly-derived MSCs were cultured in ascorbic acid supplemented medium for 14 days prior to decellularisation using two methods. 1% SDS/Triton X-100 (ST) or 20?mM ammonia/Triton X-100 (AT). CCs isolated from 4-week-old C57/BL6N mice were cultured on the decellularised MSC matrices, and induced to differentiate into cardiomyocytes in cardiogenic medium for 21 days. Cardiac differentiation was assessed by immunocytochemistry and qPCR. All data were analysed using ANOVA. Results In?vitro decellularisation using ST method caused matrix delamination from the wells. In contrast, decellularisation using AT improved the matrix retention up to 30% (p??0.05). This effect was further enhanced when MSCs were cultured in cardiogenic medium, with a matrix retention rate up to 90%. CCs cultured on cardiogenic MSC matrix (ECMsupcardio/sup), however, did not significantly improve its proliferation after 3 days (p??0.05), but the viability of CCs was augmented to 67.2?±?0.7% after 24-h exposure to Hsub2/subOsub2/sub stress as compared to 42.9?±?0.5% in control CCs ( p ??0.05). Furthermore, CCs cultured on cardiogenic MSC matrices showed 1.7-fold up-regulation in cardiac troponin I (cTnI) gene expression after 21 days (p??0.05). Conclusion Highest matrix retention can be obtained by decellularization using Ammonia/Triton-100 in 2-D culture. ECMsupcardio/sup could rescue CCs from exogenous hydrogen peroxide and further upregulated the cardiac gene expressions, offering an alternate in?vitro priming strategy to precondition CCs which could potentially enhance its survival and function after in?vivo transplantation.
机译：目的心肌梗塞仍然是全世界头号杀手疾病。使用心脏c-kit细胞（CCs）进行的细胞治疗能够再生受损的心脏。先前的研究表明，间充质干细胞衍生的（MSC）细胞外基质可以提供结构支持，并能够调节干细胞的功能和分化。这项研究旨在评估人类MSC衍生基质对CC生长和分化的影响。方法将人沃顿氏胶冻来源的MSCs在抗坏血酸补充培养基中培养14天，然后使用两种方法脱细胞。 1％SDS / Triton X-100（ST）或20？mM氨水/ Triton X-100（AT）。在脱细胞的MSC基质上培养从4周龄的C57 / BL6N小鼠中分离的CC，并诱导其在心源性培养基中分化为心肌细胞21天。通过免疫细胞化学和qPCR评估心脏分化。使用ANOVA分析所有数据。结果使用ST方法进行的体外脱细胞导致孔中基质分层。相反，使用AT的脱细胞作用将基质保留提高了30％（p≤＜0.05）。当在心源性培养基中培养MSC时，基质保留率高达90％，从而进一步增强了这种作用。但是，在心源性MSC基质（ECM cardio ）上培养的CCs在3天后并没有显着改善其增殖（p？<？0.05），但是CCs的活力增加到了67.2？±？0.7。暴露于H 2 O 2 应激24小时后的百分率，相比之下，对照CC为42.9％±0.5％（p <0.05）。另外，在心源性MSC基质上培养的CC在21天后显示出心肌肌钙蛋白I（cTnI）基因表达的1.7倍上调（p≤0.05）。结论使用氨水/ Triton-100在二维培养物中脱细胞可以获得最高的基质保留率。 ECM cardio 可以从外源性过氧化氢中拯救CC，并进一步上调心脏基因的表达，从而提供了一种替代的体外启动策略来预处理CC，这可能会提高其在体内移植后的存活率和功能。

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