P–102 GM-CSF (granulocyte-macrophage colony-stimulating factor) as a sperm medium supplement improves sperm quality in Oligoastenoteratospermia (OAT) men by activating the PI3K/Akt pathway

Abstract

Abstract Study question Can GM-CSF as a sperm medium supplement improve sperm quality in OAT men? Summary answer GM-CSF can be used to improve sperm quality in OAT men by activating the PI3K/Akt pathway. What is known already OAT patients have very low sperm parameters, including morphology, count, and motility, which can adversely affect the results of assisted reproduction technologies. It seems the development of sperm media is necessary to improve the sperm parameters of these patients. GM-CSF is a natural growth factor produced by the reproductive organs, culture media supplemented with GM-CSF is widely commercially available and enhances the embryo development and implantation rate. Studies show that this growth factor in the semen of infertile men is lower than that of fertile men. However, there is no study to assess the effect of GM-CSF on sperm quality. Study design, size, duration In the present study, Semen specimens were collected from 20 OAT patients who have male infertility factors, according to WHO criteria. After the swim-up washing procedure, each of the samples is divided into two groups; experiment, and control. In the experimental group, samples are incubated with a medium containing 2 ng/ml GM-CSF for one hour, yet, in the control group, the sperms are incubated without GM-CSF for the same time. Participants/materials, setting, methods The sperm motility was examined with phase-contrast microscopy, Eosin-nigrosin staining method was used to assess sperm viability. The expression of sperm glucose transporters (GLUT 1, 3) was determined using immunofluorescent staining, the ratio of pAkt to total Akt was assessed by the Western blotting method. Image J software was used to quantify results; the data was analyzed by SPSS software. P-value<0.05 was considered statistically significant. Main results and the role of chance As compared to the control group, supplementation with GM-CSF improved sperm progressive motility, enhanced GLUT 1 and 3, and p-AKT/AKT expression (P < 0.05). There was no significant difference between the viability of the control and experimental groups. Limitations, reasons for caution This study needs further investigations, including Real-time PCR for the genes of this signaling pathway which is ongoing. Wider implications of the findings: We showed for the first time that GM-CSF can improve sperm quality by influencing motility and energy metabolism in spermatozoa which can be affected by increasing the phosphorylation of AKT. This growth factor could be an appropriate supplement in sperm media for OAT patients. Trial registration number IRCT20200519047508N1