Hormonal induction of all stages of spermatogenesis in germ‐somatic cell coculture from immature Japanese eel testis
Hormonal induction of all stages of spermatogenesis in germ‐somatic cell coculture from immature Japanese eel testis
复制标题
DOI：
10.1046/j.1440-169x.1996.t01-2-00004.x
复制
发表时间：
1996-06
期刊：
影响因子：
4.6
通讯作者：
C. Miura;T. Miura;M. Yamashita;K. Yamauchi;Y. Nagahama
中科院分区：
文献类型：
--
作者：
C. Miura;T. Miura;M. Yamashita;K. Yamauchi;Y. Nagahama
在培育的雄性鳗鱼中，精原细胞是唯一存在于睾丸中的生殖细胞。我们以前使用器官培养系统的研究表明，促性腺激素和11-酮睾酮(11-KT，硬骨鱼中的一种有效雄激素)在体外可以诱导精子发生的所有阶段。为了深入研究11-KT体外诱导精子发生的调控机制，特别是生殖细胞和睾丸体细胞之间的相互作用，我们建立了一种新的培养体系，即生殖细胞和体细胞通过离心法聚集成颗粒后进行共培养。从未成熟的鳗鱼睾丸中分离出生殖细胞(精原细胞)和体细胞(主要是支持细胞)。分离的生殖细胞和体细胞共培养没有形成聚集，即使在11-KT存在的情况下也不能诱导精子发生。相反，将分离的生殖细胞和体细胞通过离心法制成颗粒，然后与11-KT培养30天，则诱导了从有丝分裂前精原细胞到精子的整个精子发生过程。然而，在没有11-KT的情况下，即使生殖细胞和体细胞聚集在一起，也不能诱导精子发生。这些结果表明，生殖细胞与支持细胞的物理接触是诱导精子发生对11-KT的反应所必需的。
In cultivated male eel, spermatogonia are the only germ cells present in testis. Our previous studies using an organ culture system have shown that gonadotropin and 11‐ketotestosterone (11‐KT, a potent androgen in teleost fishes) can induce all stages of spermatogenesis in vitro. for detailed investigation of the control mechanisms of spermatogenesis, especially of the interaction between germ cells and testicular somatic cells during 11‐KT‐induced spermatogenesis in vitro, we have established a new culture system in which germ cells and somatic cells are cocultured after they are aggregated into pellets by centrifugation. Germ cells (spermatogonia) and somatic cells (mainly Sertoli cells) were isolated from immature eel testis. Coculture of the isolated germ cells and somatic cells without forming aggregation did not induce spermatogenesis, even in the presence of 11‐KT. In contrast, when isolated germ cells and somatic cells were formed into pellets by centrifugation and were then cultured with 11‐KT for 30 days, the entire process of spermatogenesis from premitotic spermatogonia to spermatozoa was induced. However, in the absence of 11‐KT in the culture medium spermatogenesis was not induced, even when germ cell and somatic cells were aggregated. These results demonstrate that physical contact of germ cells to Sertoli cells is required for inducing spermatogenesis in response to 11‐KT.