@Article{ecn.2023.0490,
AUTHOR = {Areej Jorban, Eitan Lunenfeld, Mahmoud Huleihel},
TITLE = {Granulocyte-macrophage colony-stimulating factor (GM-CSF)-induced  maturation of spermatogonial cells from prepubertal mice <i>in vitro</i> is enhanced  by testosterone},
JOURNAL = {European Cytokine Network},
VOLUME = {34},
YEAR = {2023},
NUMBER = {4},
PAGES = {54--62},
URL = {http://www.techscience.com/ECN/v34n4/64884},
ISSN = {1952-4005},
ABSTRACT = {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 <i>in vitro</i>. We isolated cells from seminiferous tubule cells of seven-day-old mice and cultured them <i>in vitro</i> 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 <i>in vitro</i>. 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 <i>in vitro</i> and in <i>in vivo</i>.},
DOI = {10.1684/ ecn.2023.0490}
}