Institute of Medical Sciences, Foresterhill, University of Aberdeen, Aberdeen, UK.
Institute of Ophthalmology, University College London, London, UK.
Commun Biol. 2021 Jun 28;4(1):802. doi: 10.1038/s42003-021-02322-8.
Regeneration of the testis from pluripotent stem cells is a real challenge, reflecting the complexity of the interaction of germ cells and somatic cells. Here we report the generation of testicular somatic cell-like cells (TesLCs) including Sertoli cell-like cells (SCLCs) from mouse embryonic stem cells (ESCs) in xeno-free culture. We find that Nr5a1/SF1 is critical for interaction between SCLCs and PGCLCs. Intriguingly, co-culture of TesLCs with epiblast-like cells (EpiLCs), rather than PGCLCs, results in self-organised aggregates, or testicular organoids. In the organoid, EpiLCs differentiate into PGCLCs or gonocyte-like cells that are enclosed within a seminiferous tubule-like structure composed of SCLCs. Furthermore, conditioned medium prepared from TesLCs has a robust inducible activity to differentiate EpiLCs into PGCLCs. Our results demonstrate conditions for in vitro reconstitution of a testicular environment from ESCs and provide further insights into the generation of sperm entirely in xeno-free culture.
多能干细胞再生睾丸是一个真正的挑战，反映了生殖细胞和体细胞相互作用的复杂性。在这里，我们报告了在无饲养层培养条件下，从小鼠胚胎干细胞（ESCs）中生成包括支持细胞样细胞（SCLCs）在内的睾丸体细胞样细胞（TesLCs）。我们发现 Nr5a1/SF1 对于 SCLCs 和 PGCLCs 之间的相互作用至关重要。有趣的是，TesLCs 与类上胚层细胞（EpiLCs）而不是 PGCLCs 的共培养导致自我组织的聚集物或睾丸类器官的形成。在类器官中，EpiLCs 分化为 PGCLCs 或类精原细胞，它们被包含在由 SCLCs 组成的类似于生精小管的结构中。此外，TesLCs 制备的条件培养基具有很强的诱导活性，可将 EpiLCs 分化为 PGCLCs。我们的结果证明了从 ESCs 体外重建睾丸环境的条件，并为在无饲养层培养条件下完全生成精子提供了进一步的见解。
