Bourdon V, Lablack A, Abbe P, Segretain D, Pointis G
Groupe de Recherche sur l'Interaction Gamétique, CJF INSERM 95/04, Faculté de Médecine, Nice, France.
Biol Reprod. 1998 Feb;58(2):591-9. doi: 10.1095/biolreprod58.2.591.
In the present study we report the isolation and characterization of a clonal Sertoli cell line (42GPA9) from sexually mature polyoma virus large T (PyLT) transgenic mice. The cells multiplied indefinitely and expressed large T antigen. The 42GPA9 cell line expressed biochemical features associated with normal Sertoli cells. Transferrin, sulfated glycoprotein-2, and the ligand of c-kit were detected by reverse transcription-polymerase chain reactions and Western blot analyses. Zymographic analysis indicated that the 42GPA9 cell line secreted tissue-type plasminogen activator. These cells also retained FSH receptors as suggested by their specific responsiveness to the gonadotropin (morphological and phagocytic changes, stimulation of cAMP production) and the detection of FSH receptor mRNAs. Another original aspect of the 42GPA9 cell line is its ability to form tight junctions at confluency as demonstrated by electron microscopic study and immunolocalization of the tight junction-associated protein zonula occludens 1. The 42GPA9 cell line, which has retained several important hallmarks of normal Sertoli cells, may prove useful for further studies on Sertoli cell behavior and on Sertoli-germ cell interactions in the mature testis.
在本研究中，我们报告了从性成熟的多瘤病毒大T（PyLT）转基因小鼠中分离并鉴定出一种克隆性支持细胞系（42GPA9）。这些细胞能无限增殖并表达大T抗原。42GPA9细胞系表现出与正常支持细胞相关的生化特征。通过逆转录-聚合酶链反应和蛋白质免疫印迹分析检测到了转铁蛋白、硫酸化糖蛋白-2和c-kit配体。酶谱分析表明42GPA9细胞系分泌组织型纤溶酶原激活剂。这些细胞对促性腺激素具有特异性反应（形态学和吞噬作用改变、刺激cAMP产生）以及检测到促卵泡激素（FSH）受体mRNA，提示它们保留了FSH受体。42GPA9细胞系的另一个独特之处在于，电子显微镜研究和紧密连接相关蛋白闭合蛋白1的免疫定位表明，其在汇合时能够形成紧密连接。42GPA9细胞系保留了正常支持细胞的几个重要特征，可能对进一步研究成熟睾丸中支持细胞的行为以及支持细胞与生殖细胞的相互作用有用。
