The Shraga Segal Dept. of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel, The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel.
Adelson School of Medicine, Ariel University, Ariel, Israel.
Eur Cytokine Netw. 2023 Dec 1;34(4):54-62. doi: 10.1684/ecn.2023.0490.
Spermatogenesis is the complicated process of sperm generation. During this process, spermatogonial cells proliferate and differentiate via meiotic and post-meiotic stages to produce mature sperm. This process is under the regulation of testicular autocrine/paracrine factors. In addition, endocrine factors are crucial to complete spermatogenesis. We aimed to localize granulocyte-macrophage colony-stimulating factor (GM-CSF) and its receptor (GM-CSFR) in testicular cells and further evaluate its involvement in the development of spermatogenesis in vitro. We isolated cells from seminiferous tubule cells of seven-day-old mice and cultured them in vitro using a methylcellulose culture system (MCS), in the presence of GM-CSF and/or testosterone for four weeks. The cells were then examined for markers of different stages of spermatogenesis by immunofluorescence staining and/or qPCR analyses. Our results revealed the presence of GM-CSF and GM-CSFR in testicular cells (premeiotic and meiotic cells as well as somatic cells; Leydig and Sertoli cells). We further demonstrated the development of colonies/spheroids in the MCS which contained pre-meiotic, meiotic, and post-meiotic cells. The addition of GM-CSF to the MCS significantly increased the percentage of pre-meiotic and meiotic cells compared to control. Furthermore, the addition of GM-CSF and testosterone together significantly increased the percentage of cells in the post-meiotic stage compared to the addition of each separately. In conclusion, our results indicate that testicular cells express GM-CSF/GM-CSFR, and that GM-CSF is involved in the development of different stages of spermatogenesis in vitro. Furthermore, testosterone enhances the development of spermatogenic cells and potentiates the effect of GMCSF on the development of post-meiotic cells. These findings provide evidence that GM-CSF and testosterone are involved in the development of spermatogenesis in vitro and in vivo. In brief: Testicular somatic and germ cells express GM-CSF and GM-CSFR. Our study suggests that testicular GM-CSF is involved in the development of spermatogenesis, which is potentiated by testosterone.
精子发生是精子生成的复杂过程。在此过程中，精原细胞通过减数分裂和减数分裂后阶段增殖和分化，产生成熟精子。这个过程受睾丸自分泌/旁分泌因子的调节。此外，内分泌因素对完成精子发生至关重要。我们的目的是定位粒细胞-巨噬细胞集落刺激因子 (GM-CSF) 和其受体 (GM-CSFR) 在睾丸细胞中，并进一步评估其在体外精子发生发育中的作用。我们从 7 天大的小鼠的曲细精管细胞中分离细胞，并在含有 GM-CSF 和/或睾酮的情况下使用甲基纤维素培养系统 (MCS) 在体外培养 4 周。然后通过免疫荧光染色和/或 qPCR 分析检查细胞不同阶段的精子发生标志物。我们的结果表明 GM-CSF 和 GM-CSFR 存在于睾丸细胞中（减数分裂前和减数分裂细胞以及体细胞；Leydig 和 Sertoli 细胞）。我们进一步证明了 MCS 中包含减数分裂前、减数分裂和减数分裂后细胞的集落/球体的发育。与对照组相比，GM-CSF 添加到 MCS 中显著增加了减数分裂前和减数分裂细胞的百分比。此外，与单独添加每个细胞相比，GM-CSF 和睾酮的共同添加显著增加了减数分裂后细胞的百分比。总之，我们的结果表明睾丸细胞表达 GM-CSF/GM-CSFR，GM-CSF 参与体外不同阶段的精子发生发育。此外，睾酮增强了生殖细胞的发育，并增强了 GMCSF 对减数分裂后细胞发育的作用。这些发现为 GM-CSF 和睾酮参与体内外精子发生发育提供了证据。简而言之：睾丸体细胞和生殖细胞表达 GM-CSF 和 GM-CSFR。我们的研究表明，睾丸 GM-CSF 参与精子发生的发育，这是由睾酮增强的。
