Tres L L, Mesrobian H G, Abdullah M, Kierszenbaum A L
Department of Pediatrics, School of Medicine, University of North Carolina, Chapel Hill.
J Urol. 1989 Apr;141(4):1003-9. doi: 10.1016/s0022-5347(17)41086-x.
A procedure is described for the preparation and maintenance of human Sertoli-spermatogenic cell cocultures using biopsies of normal and undescended testis. The evaluation of cell viability and differentiation potential of cultured spermatogenic cell was monitored by [3H]thymidine labeling combined with light microscopic autoradiography. Spermatogenic cells of the same progeny, connected by intercellular bridges, display synchronous DNA synthesis when labeled at the preleptotene stage of meiotic prophase. The pattern of [35S]methionine-labeled secretory proteins was determined by two-dimensional electrophoresis and autoradiography during testicular development and compared with these observed in human Sertoli-spermatogenic cell cocultures prepared from same specimens. Both testicular tissue and cocultured Sertoli and spermatogenic cells displayed comparable patterns of secretory proteins. A discrete group of acidic polypeptides of Sertoli cell origin enhanced their radiolabeling intensity during testicular development. Results of this paper indicate that human Sertoli-spermatogenic cell cocultures could be valuable for assessing the proliferation and differentiation potential of spermatogenic cells in children with cryptorchid testis.
本文描述了一种使用正常和隐睾活检组织制备和维持人支持细胞-生精细胞共培养物的方法。通过[3H]胸腺嘧啶核苷标记结合光学显微镜放射自显影术监测培养的生精细胞的细胞活力和分化潜能评估。处于减数分裂前期细线期的同一后代的生精细胞，通过细胞间桥相连，在标记时显示同步DNA合成。在睾丸发育过程中，通过二维电泳和放射自显影术确定[35S]甲硫氨酸标记的分泌蛋白模式，并与从相同标本制备的人支持细胞-生精细胞共培养物中观察到的模式进行比较。睾丸组织以及共培养的支持细胞和生精细胞均显示出可比的分泌蛋白模式。一组离散的支持细胞来源的酸性多肽在睾丸发育过程中增强了其放射性标记强度。本文结果表明，人支持细胞-生精细胞共培养物对于评估隐睾患儿生精细胞的增殖和分化潜能可能具有重要价值。
