In mouse fetal gonads, sex differentiation begins at 10.5-11.5 days postcoitum (dpc). With XY gonads of 12.5 dpc, cord-like structures are visible and stromal cells migrate from adjacent mesonephros, unlike in XX gonads. However, the migrated mesonephric cells, except for the endothelial cells, have not been specifically identified because they have not expressed differentiation markers over the course of organ coculture in previous experiments. In this study, we have for the first time succeeded in isolating only the mesonephric cells that migrate into the XY gonad from the mesonephros with alive and then cultured these cells in vitro through the use of an organ coculture system using EGFP-transgenic mice and a FACS Vantage. The migrated and isolated cells were used for morphological and molecular characterization. The migrated mesonephric cells contained three cell forms; a sharp cell form, a round cell form, and a cluster-forming cell. The sharp cells have the characters of peritubular myoid cells. The round cells and cluster-forming cells have the potential to differentiate into Leydig cells, as some of them are 3beta-HSD-positive. In in vitro culture of migrated mesonephric cells, the cluster-forming cells proliferated well and then differentiated into round cells, suggesting that the cluster-forming cells may be stem or precursor cells for the round cells. Thus, our findings provide important information related to the migration and differentiation of migrated mesonephric cells in the XY gonad.

译文

在小鼠胎儿性腺中,性别分化开始于coitum后10.5-11.5天 (dpc)。与12.5 dpc的XY性腺不同,脐带样结构是可见的,并且基质细胞从相邻的中肾迁移,这与XX性腺不同。但是,除内皮细胞外,迁移的中肾细胞尚未得到明确鉴定,因为它们在先前的实验中在器官共培养过程中没有表达分化标记。在这项研究中,我们首次成功地仅从中肾存活的中肾细胞中分离出了迁移到XY性腺中的中肾细胞,然后通过使用EGFP转基因小鼠的器官共培养系统在体外培养这些细胞和FACS优势。迁移和分离的细胞用于形态和分子表征。迁移的中肾细胞包含三种细胞形式; 尖锐的细胞形式,圆形细胞形式和簇形成细胞。尖锐细胞具有肾小管周围肌样细胞的特征。圆形细胞和簇形成细胞具有分化为Leydig细胞的潜力,因为其中一些细胞是3β-hsd阳性。在迁移的中肾细胞的体外培养中,簇形成细胞增殖良好,然后分化为圆形细胞,这表明簇形成细胞可能是圆形细胞的干细胞或前体细胞。因此,我们的发现提供了与XY性腺中迁移的中肾细胞的迁移和分化有关的重要信息。

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