Pluripotent stem cells (PSCs), including induced PSCs, hold great potential for personalized disease modeling, drug testing and cell-based therapeutics. However, cells differentiated from PSCs remain immature in a dish, and thus there are serious caveats to their use in modeling adult-onset diseases such as cardiomyopathies and Alzheimer's disease. By taking advantage of knowledge gained about mammalian development and from bioinformatics analyses, we recently developed a neonatal rat system that enables maturation of PSC-derived cardiomyocytes into cardiomyocytes analogous to those seen in adult animals. Here we describe a detailed protocol that describes how to initiate the in vitro differentiation of mouse and human PSCs into cardiac progenitor cells, followed by intramyocardial delivery of the progenitor cells into neonatal rat hearts, in vivo incubation and analysis. The entire process takes ∼6 weeks, and the resulting cardiomyocytes can be analyzed for morphology, function and gene expression. The neonatal system provides a valuable tool for understanding the maturation and pathogenesis of adult human heart muscle cells, and this concept may be expanded to maturing other PSC-derived cell types, including those containing mutations that lead to the development of diseases in the adult.

中文翻译：

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将新生大鼠系统用作生物培养箱，以从人和小鼠多能干细胞生成成年状的成熟心肌细胞。

多能干细胞（PSC）包括诱导的PSC，在个性化疾病建模，药物测试和基于细胞的治疗方法方面具有巨大潜力。但是，从PSC分化出来的细胞在培养皿中仍未成熟，因此在模拟成人病如心肌病和阿尔茨海默氏病方面存在严重的警告。通过利用从哺乳动物发育中获得的知识以及从生物信息学分析中获得的知识，我们最近开发了一种新生大鼠系统，该系统可使PSC衍生的心肌细胞成熟成类似于成年动物中看到的心肌细胞。在这里，我们描述了一个详细的协议，该协议描述了如何启动小鼠和人PSC体外分化为心脏祖细胞，然后将祖细胞在心肌内递送到新生大鼠心脏，体内孵育和分析。整个过程大约需要6周的时间，并且可以分析所得的心肌细胞的形态，功能和基因表达。新生儿系统为了解成人人心肌细胞的成熟和发病机理提供了有价值的工具，并且该概念可能会扩展到成熟其他PSC衍生的细胞类型，包括那些导致成人疾病发展的突变细胞。