分离的大鼠睾丸曲细精管体外重组的观察

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。

OBSERVATIONS ON THE REORGANIZATION OF THE SEMINIFEROUS TUBULES OF THE RAT TESTIS IN VITRO

The Sertoli cell-rich or Sertoli-germ cell aggregates of 15-21 day immature rat testis were cultured in serum-free Ham F-12 medium for observation of the reorganization of the seminiferous tubules in vitro. The results showed that after first week of culture, the cell aggregates were spreaded on the bottom of culture dish as a monolayer consisted of Sertoli cells as well as spermatogenic cells. While after second week these monolayer cell cultures rearranged and transformed into cellular cords which connected each other to form a cellular rete. During this time, there were many small cells with long cytoplasmic processes appeared in the cultures. They looked like immature spermatozoon-like cells which were released from the cellular cords and floated in the medium, however no movement was detected. After third week, the cellular cords developed into a solid or tubule-like structures consisted of Sertoli cells and spermatogenie cells in different stages of spermatogenesis.These cultures have been studied by phase-microscopy in vitro, light and electron microscopy on semithin and ultrathin sections. These studies revealed that the Sertoli cells and Sertoli-germ cell aggregates of immature rat testis, in vitro, not only developed into monolayer cell culture as mentioned, but were also able to be further reconstructed, or reorganized, in some extent, to solid and tubule-like structures. The possible significance and mechanism were discussed.