Permeable collagen-glycosaminoglycan cross-linked copolymers for the study of biological responses of cocultured Sertoli and spermatogenic cells.
Permeable collagen-glycosaminoglycan cross-linked copolymers for the study of biological responses of cocultured Sertoli and spermatogenic cells.
复制标题
可渗透的胶原蛋白-糖胺聚糖交联共聚物，用于研究共培养的支持细胞和生精细胞的生物反应。
DOI：
10.1007/bf00130512
复制
发表时间：
1992
影响因子：
6.1
通讯作者：
Kierszenbaum,AL
中科院分区：
文献类型：
--
作者：
Tres,LL;Cahn,F;Kierszenbaum,AL
开发了一种新型的胶原-糖胺聚糖(C-GAG)底物，以克服HATF硝基纤维素底物的光学不透明度，为支持细胞和生精细胞的共培养提供更具生理渗透性的底物。共培养物制备在光学透明的C-GAG圆盘上，附着在聚酯网状物上，以便于处理。支持细胞呈立方-柱状，接种后12d，大量精原细胞和初级精母细胞通过细胞间桥连接在支持细胞的基外面和顶面。采用大鼠支持-生精细胞共培养方法，研究毒物对大鼠体外生精的影响。在我们最初的研究中，我们测试了有毒棉酚对与Sertoli细胞在非渗透(塑料)和可渗透底物(Hatf硝酸纤维素)上共培养的生精细胞的影响，在获得连续的电阻细胞单层后，在两种标准培养条件下和灌流期间。在生精细胞(精原细胞和精母细胞)中观察到选择性线粒体结构损伤，而在共存的Sertoli细胞中未观察到。这种损伤是时间依赖的(15-60分钟)和剂量依赖的(0.1-10微米)，在灌流条件下发展得更快。据报道，在完整的动物中也有类似的线粒体损伤，但需要更高的浓度(Mg)和更长的给药时间(月)才能检测到。研究正在评估在不同类别的C-GAG底物上制备的共培养中，额外的有毒化学物质对支持细胞和生精细胞功能特性的影响。
A novel collagen-glycosaminoglycan (C-GAG) substrate was developed to overcome the optical opacity of a HATF nitrocellulose substrate and to provide a more physiological permeable substrate for cocultured Sertoli and spermatogenic cells. Cocultures were prepared on optically transparent C-GAG discs attached to a polyester mesh to facilitate handling. Sertoli cells displayed a cuboidal-to-columnar shape; a large number of spermatogonia and primary spermatocytes connected by intercellular bridges were associated with basolateral and apical surfaces of Sertoli cells up to 12 days after plating. Rat Sertoli-spermatogenic cell cocultures have been used for testing the effect of toxicants on rat spermatogenesisin vitro. In our initial studies, we tested the effects of the toxicant gossypol on spermatogenic cells cocultured with Sertoli cells on nonpermeable (plastic) and permeable substrates (HATF nitrocellulose) under both standard culture conditions and during perifusion after achieving a continuous electrical-resistant cell monolayer. A selective mitochondrial structural damage was observed in spermatogenic cells (spermatogonia and spermatocytes) but not in the coexisting Sertoli cells. This damage was time- (15–60 min) and dose-dependent (0.1–10µM) and developed more rapidly under perifusion conditions. Similar mitochondrial damage was reported in the intact animal but required higher concentrations (mg) and longer administration time (months) for detection. Studies are in progress to evaluate the effect of additional toxic chemical agents on functional properties of Sertoli and spermatogenic cells in cocultures prepared on various classes of C-GAG substrates.