Tres L L, Kierszenbaum A L
Proc Natl Acad Sci U S A. 1983 Jun;80(11):3377-81. doi: 10.1073/pnas.80.11.3377.
Spermatogenic cells from 20- to 35-day-old rats were grown in vitro in the presence of Sertoli cells maintained in serum-free hormone/growth factor-supplemented medium and alternating high/low concentrations of follicle-stimulating hormone in the medium. In cell reaggregation experiments, spermatogenic cells reassociate with Sertoli cells but not with peritubular cells or cell-free substrate. Autoradiographic experiments using [3H]thymidine as a labeled precursor for DNA synthesis show that spermatogonia and preleptotene spermatocytes, connected by cytoplasmic bridges, have a synchronous S phase. [3H]Thymidine-labeled preleptotene spermatocytes progress until later stages of meiotic prophase. Time-lapse cinematographic studies of Sertoli/spermatogenic cell cocultures show three major movement patterns. While Sertoli cell cytoplasmic processes between adjacent cells display tensional forces, spermatogonia are engaged in oscillatory cell movements different from the nuclear rotation observed in meiotic prophase spermatocytes. Results of this study show that the proliferation of premeiotic cells and the differentiation of meiotic prophase cells do occur in vitro in association with Sertoli cells maintained in a medium that allows differentiated cell functions.
将20至35日龄大鼠的生精细胞与支持细胞共同培养于体外，支持细胞维持在无血清且添加了激素/生长因子的培养基中，并在培养基中交替使用高/低浓度的促卵泡激素。在细胞重聚集实验中，生精细胞与支持细胞重新结合，但不与睾丸周细胞或无细胞底物结合。使用[3H]胸腺嘧啶核苷作为DNA合成的标记前体的放射自显影实验表明，通过细胞质桥相连的精原细胞和前细线期精母细胞具有同步的S期。[3H]胸腺嘧啶核苷标记的前细线期精母细胞会发育至减数分裂前期的后期阶段。对支持细胞/生精细胞共培养物进行的延时电影摄影研究显示出三种主要的运动模式。相邻细胞之间的支持细胞细胞质突起表现出张力，而精原细胞则进行振荡性细胞运动，这与减数分裂前期精母细胞中观察到的核旋转不同。本研究结果表明，减数分裂前细胞的增殖以及减数分裂前期细胞的分化确实在体外与维持在允许分化细胞功能的培养基中的支持细胞相关联地发生。
