Study on the Th 17 Cytokines Expression in Human Endometrium and Implantation Using a Microfluidic Capillary-Endometrial Cell Culture Chip

The molecular events of embryonic attachment to the endometrial epithelium and subsequent invasion into the stroma have long been of interest, scientifically to reproductive biologists and clinically to couples with infertility and to the physicians caring for them. Understanding the molecular factors involved in each phase of the implantation process is critical to comprehending the mechanisms, which control reproduction. Recently, cytokines have become increasingly implicated in embryo implantation. Intercellular interaction during embryo implantation requires a molecular dialogue between both endometrium and embryo. Th2 cytokines may play an important role in the maintenance of murine pregnancy by inhibiting Th1 responses that induce pregnancy failure. The immune system working in the endometrium could be modulated by various locally produced cytokines, such as TNF, IL-1, IL-16, IL-18 and IL-15. Recently, the existence of possible roles for the TH 17 cytokines throughout the human reproductive system has been amply documented. This hypothesis has since been adopted in the maintenance of normal human pregnancy and pregnancy failure. The Th1 ⁄ Th2 paradigm has recently been reconstituted to include a third population of T helper cells that produce IL-17, which are designated as Th17 cells. Th17 cells have specific roles in host defense against extracellular bacteria and fungi and play an important role in the induction of autoimmune diseases. In addition, IL-21 is an autocrine cytokine that functions as a positive feedback signal for the Th17 cell. However, the IL-21R is expressed not only on Th17 cells, but on a variety of cells such as B and natural killer (NK) cells and dendritic cells (DCs), resulting in pleiotropic effects of IL-21. For IL-22 the same story applies: publications vary from IL-22 being pathogenic to IL-22 as a protective factor. Microfluidic devices are being developed for biological assays with the potential to perform high throughput analysis, utilize small amounts of samples/reagents, and for single cell analysis. Therefore, we have developed a microfluidic chip that mimics the blood vessel microenvironment. Microfluidic chip was designed by sandwiching a micro pore array silicon chip between two polydimethylsiloxane (PDMS) chips, To further investigate the role of TH 17 cytokine system during the interaction of embryo and maternal uterine endometrium communication and regulation through cytokine network, we intend to conduct the follow studies: 1). To examine human endometrium obtained from hysterectomy specimen to determine TH 17 cytokine system mRNA expression by PCR. 2). To further investigate quantitative amount of human endometrial cell TH 17 cytokine system at different menstrual cycle (proliferative and luteal phase) mRNA expression by QC PCR. 3). To further examine human endometrium to determine TH 17 cytokine system protein expression by immunohistochemistry. 4). To further establish the balance between the TH17/TH1/TH2 endometrial cell in different menstrual cycle 5). To establish the human endometrial stromal cells and trophoblast cell culture model for study of embryo implantation. 6). To establish PDMS & silicon based microfluidic capillary-endometrial cell culture chip used for endometrial cell culture and trophoblast invasion.

Project ID:PC10107-0352
External Project ID:NSC101-2314-B182A-133