Effect of granulocyte colony-stimulating factor on vascular endothelial growth factor expression in human endometrial cell culture

Background: A clinical observation of the results of treatment with recombinant granulocyte colony-stimulating factor (G-CSF) in patients with idiopathic neutropenia, a condition characterized by infertility and miscarriage, demonstrated spontaneous pregnancy in patients with a history of infertility. These results served as the basis for the use of G-CSF in refractory endometrial syndrome.

Aim: The work aimed to investigate the effect of G-CSF on the expression of the vascular endothelial growth factor (VEGF) gene and the transcription factor HOXA10 in primary cultures of human endometrial cells.

Methods: The study included eight female patients who were examined before enrolling in assisted reproductive technology programs. Endometrial aspiration biopsy was performed on days 5–10 of the menstrual cycle. The samples were cultured for 72 hours in DMEM supplemented with various concentrations of G-CSF (1 and 100 mg/mL). The expression of VEGF and HOXA10 genes was assessed using real-time polymerase chain reaction, and VEGF concentration in the culture medium was determined by enzyme-linked immunosorbent assay. Statistical analysis was performed using one-way ANOVA followed by Tukey’s test.

Results: A dose-dependent stimulatory effect of G-CSF on VEGF [F(2, 21) = 31.55; p < 0.001] and HOXA10 [F(2, 21) = 14.31; p < 0.001] gene expression was established. Upregulation was observed already at 1 mg/mL, whereas significant differences were achieved at 100 mg/mL. A significant increase in VEGF concentration in the culture medium was also detected in response to G-CSF [F(2, 21) = 109.4; p < 0.001]. The stimulatory effect was observed in all examined endometrial samples.

Conclusion: G-CSF exerts pleiotropic effects by stimulating VEGF and HOXA10 expression in endometrial cells in vitro. These mechanisms may underlie endometrial regeneration and improved receptivity, which are of particular relevance for treating patients with refractory endometrium and recurrent implantation failure in assisted reproductive technology programs