Isolation, Characterization and Differentiation of Mouse Cardiac Progenitor Cells.
Isolation, Characterization and Differentiation of Mouse Cardiac Progenitor Cells.
复制标题
小鼠心脏祖细胞的分离、表征和分化。
DOI：
10.1007/978-1-4939-8697-2_12
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Mishra,ParasKumar
中科院分区：
文献类型：
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作者：
Yadav,SantoshKumar;Mishra,ParasKumar
尽管开发了几种策略来补充心肌梗死（MI）后死亡的心肌，干细胞治疗仍然是再生新心肌的主要方法。虽然诱导多能干细胞（iPS）和分化细胞的转分化已被用作心肌再生的新方法，但这些方法在体内研究中没有产生非常成功的心肌再生结果。新形成的心肌细胞与现有心肌细胞的异步收缩性是iPS方法的最重要问题，而转分化心肌细胞的产量非常低，并且它们以与MI心脏中现有心肌细胞相同的节律跳动的机会较少，这是转分化方法的重要警告。CSC存在于心脏中，并且它们具有分化成心肌细胞的潜力。然而，常驻社区服务中心的数量非常少。因此，重要的是在从心脏分离过程中获得CSC的最大产量。增加CSC的数量并启动其离体分化对于基于CSC的干细胞治疗至关重要。在这里，我们提出了一个更好的方法，从小鼠心脏CSCs的分离，鉴定和分化。我们还使用相差显微镜证明了在维持培养基中培养2天、3天和7天后CSC的形态学变化以及在分化培养基中培养12天的单独一组CSC。我们已经使用不同的标记物来鉴定从小鼠心脏分离的CSC，例如小鼠CSC的标记物Sca-1、心脏特异性标记物NKX 2 -5、MEF 2C、GATA 4和干性标记物OCT 4和SOX 2。为了表征分化的CSC，我们使用在分化培养基中维持12天的CSC。为了评估CSC的分化，我们测定了心肌细胞特异性标记物肌动蛋白和肌钙蛋白I的表达。总之，我们描述了一种从小鼠心脏分离、鉴定、分化和表征CSC的优雅方法。
Despite several strategies developed for replenishing the dead myocardium after myocardial infarction (MI), stem cell therapy remains the leading method to regenerate new myocardium. Although induced pluripotent stem cells (iPS) and transdifferentiation of the differentiated cells have been used as novel approaches for myocardial regeneration, these approaches did not yield very successful results for myocardial regeneration in in vivo studies. Asynchronous contractility of newly formed cardiomyocytes with the existing cardiomyocytes is the most important issue with iPS approach, while very low yield of transdifferentiated cardiomyocytes and their less chances to beat in the same rhythm as existing cardiomyocytes in the MI heart are important caveats with transdifferentiation approach. CSCs are present in the heart and they have the potential to differentiate into myocardial cells. However, the number of resident CSCs is very low. Therefore, it is important to get maximum yield of CSCs during isolation process from the heart. Increasing the number of CSCs and initiating their differentiation ex vivo are crucial for CSC-based stem cell therapy. Here, we present a better method for isolation, characterization and differentiation of CSCs from the mouse heart. We also demonstrated morphological changes in the CSCs after 2 days, 3 days, and 7 days in maintenance medium and a separate group of CSCs cultured for 12 days in differentiation medium using Phase-Contrast microscopy. We have used different markers for identification of CSCs isolated from the mouse heart such as marker for mouse CSC, Sca-1, cardiac-specific markers NKX2–5, MEF2C, GATA4, and stemness markers OCT4 and SOX2. To characterize the differentiated CSCs, we used CSCs maintained in differentiation medium for 12 days. To evaluate differentiation of CSCs, we determined the expression of cardiomyocyte-specific markers actinin and troponin I. Overall; we described an elegant method for isolation, identification, differentiation and characterization of CSCs from the mouse heart.