In vitro effect of Granulocyte-macrophage colony-stimulating factor (GM-CSF) on the expression of related genes to sperm motility and energy metabolism, and ICSI outcomes in obstructive azoospermic patients

Abstract

Abstract Background Granulocyte-macrophage colony-stimulating factor (GM-CSF) expressed in the human reproductive system, holds a pivotal role in the reproductive processes. This study investigates the in vitro effect of GM-CSF on the testicular sperm of obstructive azoospermia (OA) patients and assesses the effectiveness of GM-CSF‐supplemented sperm media in Intracytoplasmic sperm injection (ICSI) outcomes. Methods and Results Following testicular sperm extraction from 20 patients diagnosed with OA, each sample was divided into two parts: the experimental samples were incubated with the medium containing 2 ng/ml GM-CSF at 37°C for 60 min, and control samples were incubated with medium without GM-CSF. Subsequently, the oocytes retrieved from the partner were injected with sperms from treatment and the control groups. The sperm parameters ( motility, viability), the expression level of sperm motility-related genes (PIK3R1, PIK3CA, and AKT1 ), and sperm energy metabolism-related genes (GLUT1, GLUT3, and GLUT14) were assessed. Furthermore, the fertilization and cleavage rates and embryo quality were evaluated. Supplemented testicular sperm with GM-CSF significantly increased motility parameters, the mRNA expression of PIK3R1, AKT1, and GLUT3 compared to the non-treated group (p < 0.05). However, no significant differences in mRNA expression of PIK3CA, GLUT1, or GLUT14 were identified. Based on ICSI outcomes, the GM-CSF treatment group exhibited significantly higher fertilization rates (p = 0.027), cleavage rates (p = 0.001), and the proportion of good-quality embryos (p = 0.002) compared to the control group. Conclusions GM-CSF increased gene expression related to motility and energy metabolism pathway and effectively had a positive effect on the motility of testis-extracted spermatozoa and, consequently yielding positive clinical outcomes.