Silva Andreia F, Escada-Rebelo Sara, Amaral Sandra, Tavares Renata S, Schlatt Stefan, Ramalho-Santos João, Mota Paula C
Biology of Reproduction and Stem Cell Group, Center for Neuroscience and Cell Biology (CNC), University of Coimbra, Coimbra, Portugal.
Institute for Interdisciplinary Research (IIIUC), University of Coimbra, Coimbra, Portugal.
PLoS One. 2018 Feb 7;13(2):e0191912. doi: 10.1371/journal.pone.0191912. eCollection 2018.
The reduced number of animals in most wild felid populations implies a loss of genetic diversity. The death of juveniles, prior to the production of mature sperm, represents a loss of potential genetic contribution to future populations. Since 2011 mouse testicular organ culture has introduced an alternative mechanism to produce sperm in vitro from immature tissue. However, extension of this technology to other species has remained limited. We have used the domestic cat (Felis catus) as a model for wild felids to investigate spermatogenesis initiation and regulation, with the mouse serving as a control species. Testicular tissue fragments were cultured in control medium or medium supplemented with knockout serum replacement (KSR), AlbuMax, beta-estradiol or AlbuMax plus beta-estradiol. Contrary to expectations, and unlike results obtained in mouse controls, no germ cell differentiation could be detected. The only germ cells observed after six weeks of culture were spermatogonia regardless of the initial stage of tubule development in the donor tissue. Moreover, the number of spermatogonia decreased with time in culture in all media tested, especially in the medium supplemented with KSR, while AlbuMax had a slight protective effect. The combination of AlbuMax and beta-estradiol led to an increase in the area occupied by seminiferous tubules, and thus to an increase in total number of spermatogonial cells. Considering all the media combinations tested the stimulus for felid germ cell differentiation in this type of system seems to be different from the mouse. Studies using other triggers of differentiation and tissue survival factors should be performed to pursue this technology for the genetic diversity preservation in wild felids.
大多数野生猫科动物种群数量的减少意味着遗传多样性的丧失。幼年个体在产生成熟精子之前死亡,代表着对未来种群潜在遗传贡献的丧失。自2011年以来,小鼠睾丸器官培养引入了一种从未成熟组织体外产生精子的替代机制。然而,这项技术在其他物种中的应用仍然有限。我们以家猫(Felis catus)作为野生猫科动物的模型来研究精子发生的起始和调控,以小鼠作为对照物种。将睾丸组织碎片培养在对照培养基或添加了敲除血清替代物(KSR)、白蛋白(AlbuMax)、β-雌二醇或AlbuMax加β-雌二醇的培养基中。与预期相反,与在小鼠对照中获得的结果不同,未检测到生殖细胞分化。无论供体组织中肾小管发育的初始阶段如何,培养六周后观察到的唯一生殖细胞是精原细胞。此外,在所有测试培养基中,精原细胞的数量随着培养时间的延长而减少,尤其是在添加KSR的培养基中,而AlbuMax有轻微的保护作用。AlbuMax和β-雌二醇的组合导致生精小管占据的面积增加,从而使精原细胞总数增加。考虑到所有测试的培养基组合,在这种系统中,猫科动物生殖细胞分化的刺激因素似乎与小鼠不同。应该进行使用其他分化触发因素和组织存活因子的研究,以推进这项技术用于保护野生猫科动物的遗传多样性。