雄性生殖毒性的体外试验方法研究进展

Advances in the methods of evaluating male reproductive toxicity in vitro

近年来,体外替代试验在雄性生殖毒理研究中愈来愈重要,而构建稳定可靠的体外模型是其有力保障。随着生物技术的发展,体外睾丸模型已从单层的细胞培养过渡到更能模拟体内微环境的新培养体系,如微流控培养、三维(three-dimensional, 3D)培养、类器官培养等。这些培养体系不仅能模拟体内血睾屏障(blood-testis-barrier, BTB)和睾丸微环境,还能通过睾丸组织或生殖细胞培养,延长培养体系持续的时间,更为重要的是啮齿类动物、非人灵长类甚至人类生殖细胞在这些培养体系中都已成功分化出精子,这些新的培养体系也成功应用于睾丸毒物评价。然而,这些新的培养体系仍存一些局限性,如精子分化数量少、效率低以及无法完全复制体内生精进程等。本文主要综述新的睾丸体外培养体系的优缺点以及未来发展方向。

In recent years, in vitro substitution experiments have become more and more important in male reproductive toxicology research, and the construction of a stable and reliable in vitro model is a strong guarantee for the research. With the development of biotechnology, in vitro testicular models have also transitioned from a single-layer cell culture to a new culture system that can simulate in vivo microenvironment better, such as microfluidic technology, three-dimensional culture technology, and organoids, and so on. These culture systems not only can simulate the blood-testis-barrier(BTB) or in vivo testicular microenvironment, but also extend duration of the culture system through co-cultivating of testicular tissues or germ cells. More importantly, sperm has also been successfully differentiated from germ cells of rodents, non-human primate and even human beings. These new culture systems have been successfully applied to the evaluation of testicular toxicants. However, there are some limitations for them, such as small number of differentiated sperm, low efficiency and difficulty in completely simulate the process of spermatogenesis in vivo, etc. This review mainly summarizes the advantages and disadvantages of these new testis culture systems in vitro and their future development directions.