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将生后15天和20天大鼠的睾丸,去被膜后以胶原酶反复消化,使Sertoli细胞和生精细胞分离。以HamF-12无血清培养基培养子(35℃)5％CO_2温箱。经过19天的连续培养及光镜和电镜样品观察,发现分离的曲细精管上皮细胞经过贴壁生长期(培养后1～6天),形成单层培养细胞。再经索状生长期(培养后6～14天),由单层细胞变成致密的细胞索,并释放出有头尾结构的蝌蚪形精子样细胞。最后经管状生长期(培养第14天以后),细胞索发育成细胞团和类似曲细精管样的结构。光镜和电镜下见其中有管周细胞、Sertoli细胞和各级生精细胞。本文对分离的曲细精管上皮细胞在体外重组成小管样结构和体外精子发生,进行了讨论。
Sertoli cells and spermatogenic cells were isolated from the testes of rats at 15 days and 20 days after birth by repeated digestion with collagenase after they were removed. HamF-12 serum-free medium (35 ° C) 5% CO_2 incubator. After 19 days of continuous culture and light microscopy and electron microscopy samples observed isolated seminiferous tubule epithelial cells after adherent growth phase (1 to 6 days after culture), the formation of monolayer cultured cells. After the cord-like growth period (6 to 14 days after culture), the monolayer cells became dense cell lines and released tadpole-shaped sperm-like cells with a head-tail structure. Finally, during the tubular growth phase (after fourteen days of culture), the cell line develops into clusters of cells and seminiferous tubule-like structures. Light and electron microscopy, including pericytes, Sertoli cells and spermatogenic cells at all levels. In this paper, the separation of seminiferous tubule epithelial cells in vitro reorganization into tubule-like structure and in vitro spermatogenesis were discussed.