Testicular organoid models are tools to study testicular physiology, development, and spermatogenesis in vitro . However, few side-by-side comparisons of organoid generation method have been evaluated. Here, we directly tested whether the culture microenvironment is the prime determinant promoting testicular organoid self-assembly. Using Matrigel as a representative extracellular matrix (ECM), we compared multiple culture environments, 2D and 3D, ECM-free and ECM, for organoid self-assembly with immature murine testicular cells. De novo tissues were observed to self-assemble in all four culture environments tested within 72 h, however, these tissues only met requirements to be named organoids in 2D ECM and 3D ECM-free (3DF) culture methods. Based on these results, 3DF was selected for further study, and used to examine animal age as an independent variable. Organoid assembly was significantly delayed when using pubertal murine cells and entirely absent from adult mu...

中文翻译：

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睾丸类器官的形成是不成熟的体细胞的一种特性，它可以自我组装并表现出长期的激素反应性内分泌功能。

睾丸类器官模型是在体外研究睾丸生理，发育和生精的工具。但是，几乎没有评估类器官生成方法的并行比较。在这里，我们直接测试了培养微环境是否是促进睾丸类器官自组装的主要决定因素。使用基质胶作为代表性的细胞外基质（ECM），我们比较了2D和3D，无ECM和ECM的多种培养环境与未成熟鼠睾丸细胞的类器官自组装。在72小时内测试的所有四个培养环境中都观察到新生组织自组装，但是，这些组织仅满足2D ECM和无3D ECM（3DF）培养方法中被称为类器官的要求。根据这些结果，选择了3DF进行进一步研究，并将其作为独立变量来检验动物的年龄。使用青春期鼠细胞时，类器官的装配显着延迟，成年鼠完全不存在。