Research Of Intrauterine Administration Of Peripheral Blood Mononuclear Cells (PBMCs)'s Improvement On Embryonic Implantation

Objectives:To explore and discuss the impact of intrauterine administration of autologous peripheral blood mononuclear cells (PBMCs) on embryo implantation, pregnancy rate and explore its possible mechanism both clinically and experimentally, so as to explore a new treating method for patients with recurrent implantation failure (RIF).Methods:This study includes three part:Part one:animal experimental research (in vivo):Pregnant mice were randomly divided into three groups, including control group; embryo implantation dysfunction (EID) group; EID with PBMCs group. Uterine horns were excised to determine the number of pregnant mice and implantation sites on the Day7.5postcoitum. The expression levels of LIF and VEGF during the implantation window were detected with immunohistochemistry and Real Time-PCR analysis.Part two:cell experimental research (in vitro):The expression levels of IL-1? and LIF mRNA in PBMCs in the medium with or without HCG were detected by Real Time-PCR analysis. The culture medium of JAR cell were divided into four groups, including control group, HCG only group, PBMCs only group and PBMCs+HCG group. The invasion ability of JAR cells were detected by Transwell assay. And protein expression of MMP2and MMP9in JAR cell were detected by Western blotting analysis.Part three:clinical study:A prospective analysis was performed in the Medical records of173patients who had underwent IVF/ICSI-ET treatment in the Reproductive Medical Center of the Remin Hospital Wuhan University in the latest six months. Patients were divided into two groups, including control group and PBMCs treated group. General condition, fertilization and clinical pregnancy outcomes of two groups were compared.Results:Part one:Embryo implantation dysfunction model group showed a significant decrease in pregnancy rate, implantation sites and the expression of both the endometrial LIF and VEGF during the implantation window. EID with PBMCs group showed a higher pregnancy rate and endometrial LIF and VEGF expression compared to EID group.Part two:HCG can promote the expression of IL-1? and LIF mRNA in PBMCs. The number of invasive cells in PBMCs+HCG group were significantly higher than control group. And the protein expression of MMP2and MMP9in PBMCs+HCG group were significantly higher than control group.Part three:There was no significant difference in age, duration of infertility, assisted reproductive methods, number of oocytes, number of2PN, number of times before IVF, endometrial thickness in two groups. The implantation rates and clinical pregnancy rates in PBMCs treated group were significantly higher than the control group.Conclusion:1. Intrauterine administration of mouse PBMCs derived from unpregnant mice prior to embryo implant has a good influence on endometrial receptivity and embryonic implantation in EID mice.2. This study demonstrated that HCG stimulate the secretion of cytokines in human PBMCs, which could stimulates the ability of trophoblast cells to invade through the ECM in vitro, which is accompanied by increases in MMP-2and MMP-9expression.3. This preliminary clinical study in our centre showed that intrauterine administration of autologous PBMCs coculture with HCG effectively improved embryo implantation in patients with recurrent implantation failure which receiving frozen/thawed embryo transfer.