摘要
本实验应用胰酶消化后的Percoll梯度分离技术,将苗勒管上皮细胞与间质细胞分离开来,然后分别进行细胞培养。一部分与同胚龄睾丸或卵巢联合培养,观察生殖腺分泌的苗勒管抑制素(MIS)对两类细胞致死率的差异;另一部分以MIS抗体为第一抗体,HRP-羊抗兔IgG或抗生物素-胶体金为第二抗体,作免疫电镜技术,显示苗勒管上皮细胞和间质细胞上MIS受体的结合位点。实验结果表明,睾丸分泌的MIS使培养的苗勒管上皮细胞致死率与对照组相比,有非常显著差异(P<0.001),对间质细胞致死率不明显,但MIS受体结合位点的免疫反应不仅出现在苗勒管上皮细胞上,也显示在间质细胞上。本文讨论了MIS的作用机理。
An experimental study was carried out on the Mullerian duct cells, i.e. epithelial cells and stromal cells, treated with trypsin and isolated by Percoll gradient centrifugation. Then they were cultured separately. In order to observe the difference of cell death rate induced by Mullerian inhibiting substance (MIS) secreted from gonad, some cells were co-cultured with fetal testis or ovary in the same embryonic age. Other cells were used for immunocytochemistry at the electron microscopic level with anti-MIS serum as primary antibody and peroxidase labelled goat anti-rabbit IgG antiserum or streptavidin colloidal gold as secondary antibody to show the receptor localizations of MIS on epithelial cells and stromal cells. The results showed that the death rate of Mullerian duct epithelial cells induced by MIS secreted from testis was increased as compared with the control cells (P<0.001). But no statistical significant difference was noted about the stromal cells. The presence of MIS receptor localization on the plasma membrane of Mullerian duct cell, not only of the epithelial cells but also of the stromal cells, was demonstrated by indirect immunocytochemical reaction at the electron microscopic level. The mechanism of MIS action on Mullerian duct development and its physiological involution are discussed.
出处
《解剖学报》
CAS
CSCD
北大核心
1990年第3期315-319,共5页
Acta Anatomica Sinica
关键词
苗勒管
抑制素
细胞培养
免疫电镜
Mllerian duct
Mllerian inhibiting substance
Cell culture
Electron immunocytochemistry
