The effect of human embryo spent media on dendritic cells

Infertility is an increasingly common condition, with approximately every 7 th couple remaining childles s . This medical problem poses a significant burden to society and is associated with economic, social and psychological consequences. It is estimated that approximately 5% of women encounter problems w ith embryo implantation leading to recurrent implantation failure or recurrent miscarriage. While it is suspected that the maternal immune system plays a key role in implantation, the specific immunologic processes that govern implantation and embryo and fetal development remain largely unknown. Our research proposal focuses on dendritic cells (DC), which are known to increase in concentration during pregnancy. In our lab a difference was observed in medium collected from embryos that subsequently implanted compared to that of embryos that do not implant. This difference had an impact on DC. We hypothesize that factors in the medium produced by the embryo exert an influence on DC, w hic h in turn play a central role in facilitating embryo implantation and further development. With this proposal we aim to provide novel insights into the physiology of implantation. Ultimately, we hope to help women suffering from infertility to achieve successful pregnancies. As part of the in vitro fertilization process, we will obtain discarded medium from embryos and use it for the incubation of DC. We will compare the outcomes between medium from implanted and non- implanted embryos. Specifically, we will analyze and compare the morphology, function, and differentiation of DC exposed to the different mediums. We will use the methods of gene expression analysis, FACS, ELISA, IDO-Expression, co-culture of DCs with T-lymphocytes and migration experiments. In addition, the medium will be analyzed with protein arrays to test for its components, specifically cytokines and chemokines. All these methods have been established in our research laboratory. With this research project, we aim to demonstrate for the first time that embryos exert an effect on maternal dendritic cells, through which they may influence their own implantation. This immunologic- based phenomenon could play a central role in implantation and deserves further study, as suggested in this proposal. Altogether, an improved understanding of the immune systems role in implantation c ould lead to the discovery of novel methods to positively influence the immune system and thus to secure higher rates of implantation. This project is the necessary first step to working towards achieving this goal. Optimally, novel treatment options can grow out of this work, such as the establishment of a diagnos tic test for embryo implantation, which could limit the number of embryos transferred after IVF to only those with the highest implantation potential, reducing not only the chance of multiple pregnancy but also the unnecessary burden of unsuccessful implantation and/or early miscarriage.