Embryo Culture Fluid Shla-g Expression And Embryonic Development And The Planting Of Relationship

Objective:The aim of the study is to investigate the quantity and qualitation of soluble human leukocyte antigen G(sHLA-G)secreted to human embryo culture media by the embryo in cleavage stage,and to understand the relationship between Shla-g antigen presentation and embryo quality and implantation.Methods:Retrospective study involved 40 women undergoing IVF-ET between August and November 2006.All patients were＜35 years of age,and have two embryo for embryo transfer per circle.Embryos were cultured individually in 50ul droplets of culture media.On the day of transfer(D 3),20ul of supernatants were collected from each human embryo culture.A double monoclonal sandwich immunoassay kit was utilized for the quantitative measurement of sHLA-G in embryo culture supernatant,negative control was set up at the same time.160 embryo culture supernatants were assayed,and 80 out of them came from embryo transferred.Patients HLA-G+were confirmed by the patients receiving all the embryo that was positive for HLA-G,and patients HLA-G±were confirmed by the patients receiving at least one embryo that was positive for HLA-G.Otherwise,they are patients HLA-G-. Clinical pregnancy was confirmed by the presence of a fetal heart on ultrasonic examination at 6-8 weeks.The implantation rate is defined as the number of fetal hearts divided by the total number of embryos transferred.Results:In the 40 patients,there were 16 patients HLA-G+,10 patients HLA-G±and 14 patients HLA-G-.In the 16 patients HLA-G+,the pregnancy rate was 81.25%and the implantation rate was 50%.And in the 10 patients HLA-G±,the pregnancy rate was 60%and the implantation rate was 25%.In contrast,the 14 patient HLA-G-had the pregnancy rate of 35.71%and the implantation rate of 17.86%.The pregnancy rates and implantation rates were different among the three groups.Pregnancy occurred in 24 out of 40 patients,the total pregnancy rate was 60%and the implantation rate was 36.25%.There were 13 patients HLA-G+,6 patients HLA-G±and 5 patients HLA-G-in the 24 patients.Expression of HLA-G was also related to increasing cell numbers.Embryo Grade also correlated with the presence of HLA-G in embryo culture medium.The expression of sHLA-G is not different between the groupⅠ/≥6 cells and the groupⅡ/≥6 cells.Concentration of HLA-G in embryo culture media was variable and in the low range(3-10ng/ml).Conclusion:From our results,expression of HLA-G was positively correlated with the potential ability of embryo development and implantation.If day 3 embryo transfer using expression of HLA-G and embryo morphology improves IVF-ET pregnancy rate and implantation rate by increasing predictive accuracy.