van Dissel-Emiliani F M, de Boer-Brouwer M, Spek E R, van der Donk J A, de Rooij D G
Department of Veterinary Anatomy and Cell Biology, Veterinary School, Utrecht, The Netherlands.
Cell Tissue Res. 1993 Jul;273(1):141-7. doi: 10.1007/BF00304621.
Quiescent gonocytes were isolated from fetal testes of rat 18-day post coitum and cultured alone or on monolayers of somatic cells from different origins. The gonocytes specifically adhered to Sertoli cells, isolated from 21 to 23-day-old rat testes; this adherence was necessary for their survival in vitro. Addition of follicle-stimulating hormone and testosterone to these cultures did not increase the viability of the gonocytes. Serum was found to be deleterious to the germ cells. Electron-microscopic examination of Sertoli-cell-gonocyte co-cultures revealed the presence of numerous adhesion plaques between these cells, indicating that Sertoli cells and gonocytes are able to communicate in vitro. Gonocytes, in co-culture with Sertoli cells, were viable for at least 9 days. The gonocytes did not spontaneously resume proliferation. The simple culture system described in the present paper should be useful in studying the nature of the factors that are responsible for sending the quiescent gonocytes into the cell cycle and for stimulating the formation of A spermatogonia, a process characterizing the start of spermatogenesis.
静止期生殖母细胞从妊娠18天大鼠的胎儿睾丸中分离出来,单独培养或培养在来自不同来源的体细胞单层上。生殖母细胞特异性地黏附于从21至23日龄大鼠睾丸分离的支持细胞;这种黏附对于它们在体外存活是必需的。向这些培养物中添加促卵泡激素和睾酮并未提高生殖母细胞的活力。发现血清对生殖细胞有害。对支持细胞-生殖母细胞共培养物的电子显微镜检查显示这些细胞之间存在大量黏附斑,表明支持细胞和生殖母细胞在体外能够进行交流。与支持细胞共培养的生殖母细胞至少存活9天。生殖母细胞不会自发地恢复增殖。本文所述的简单培养系统应有助于研究负责使静止期生殖母细胞进入细胞周期并刺激A型精原细胞形成的因子的性质,A型精原细胞的形成是精子发生开始的一个特征性过程。