Elsenhans Anja, de Lima E Martins Lara Nathalia, Sakib Sadman, Dobrinski Ina
Department of Biochemistry and Molecular Biology, Cumming School of Medicine, and Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada.
National Research Council Canada, Ottawa, ON, Canada.
Methods Mol Biol. 2025;2954:121-134. doi: 10.1007/978-1-0716-4698-4_5.
Organoids are three-dimensional structures formed by multiple cell types that mimic the function of the respective organ in vivo. Testicular organoids are formed by testicular somatic cells and germ cells that retain tissue-specific architecture and function. Especially in male reproductive biology, testicular organoids present a platform to study cell-cell interactions, tissue development, and the germ-cell niche microenvironment. This not only holds opportunities for male fertility preservation, but it can also facilitate high-throughput drug and toxicity screening. The microwell aggregation technique presents a simple and reproducible approach for generating a large number of homogenous organoids in pyramid-shaped wells. Testes from prepubertal animals are digested, and the obtained testicular starting cell population is centrifuged into the wells and cultured to form organoids. Here, we describe our standard protocol for the generation of testicular organoids from primary rat or pig testicular cells in microwell culture, highlighting the difference between the two species and providing a strategy to enrich for germ cells.
类器官是由多种细胞类型形成的三维结构，可模拟体内相应器官的功能。睾丸类器官由睾丸体细胞和生殖细胞形成，保留了组织特异性结构和功能。特别是在男性生殖生物学中，睾丸类器官为研究细胞间相互作用、组织发育和生殖细胞生态位微环境提供了一个平台。这不仅为男性生育力保存带来了机会，还能促进高通量药物和毒性筛选。微孔聚集技术提供了一种简单且可重复的方法，用于在金字塔形孔中生成大量同质类器官。对青春期前动物的睾丸进行消化，将获得的睾丸起始细胞群体离心到孔中并培养以形成类器官。在此，我们描述了在微孔培养中从原代大鼠或猪睾丸细胞生成睾丸类器官的标准方案，强调了两种物种之间的差异，并提供了一种富集生殖细胞的策略。
