Effects of human autologous mononuclear cells on expression of leukemia inhibitory factor and its receptor in co-culture model of embryo and endometrium in vitro

Abstract

Objective To investigate the effects of human autologous mononuclear cells (PBMCs) on the expression of leukemia inhibitory factor (LIF) and leukemia inhibitory factor receptor (LIFR) on embryonic surface in co-culture model of embryo and endometrium. Methods From May 2015 to May 2016, endometrial glandular epithelial cells were isolated from 12 patients diagnosed as repeated implantation failure in the Second Affiliated Hospital of Zhengzhou University. PBMCs cells were isolated from their peripheral blood. We collected abandoned embryos (signed informed consent of abandoned embryos) from patients who had successfully delivered by assisted reproductive technology from 2009 to 2014. The study was divided into two groups. The experimental group was endometrium-embryo-PBMCs co-culture, and control group was endometrium-embryo co-culture. The day on which the culture medium containing estrogen and progesterone was added was regarded as the 0th day of culture. The concentration of LIF was detected by enzyme linked immunosorbent assay on the 1st, 3rd and 5th day after culture. The expression of LIFR on blastocyst surface was detected by immunofluorescence on the 5th and 6th day after culture in experimental group and control group. Immunofluorescence images were analyzed by Image J software. Results On the 3rd day of culture, there was no significant difference in LIF concentration between experimental group and control group (P>0.05). On the 5th day of culture, LIF concentration in experimental group ［(2 840.02±240.51) ng/L] was significantly higher than that in control group [(2 411.35±311.63) ng/L] (P=0.001). The immunofluorescence aborsance (A)value of LIFR in blastocysts of experimental group was higher than that of control group (0.255 8±0.037 2 vs. 0.190 6±0.040 4, P=0.001). Conclusion Human PBMCs can enhance the expression of LIF in endometrium and promote the expression of LIFR on blastocyst surface, which plays an important role in embryo implantation. Key words: Peripheral blood mononuclear cells; Repeated implantation failure; Human leukemia inhibitory growth factor; Co-culture