Decidual Stromal Cells Promotes Trophoblasts Migration By Inducing Matrix Metalloproteinases Expression

Objective:Trophoblasts invading into maternal endometrium is a key point to the process of pregnancy and embryo sustainable development. During embryo implantation and early pregnancy, the maternal-fetal interface is located in deciduas which are directly contacted with trophoblast. It is important for successful pregnancy that trophoblast cells migrate and invade into uterine stroma abstemiously, probably mediated by degradation of extracellular matrix which requires a balance between MMPs and TIMPs. The migration and invasion ability of trophoblastic cells are closely related with their surrounding environment, upon which uterine deciduas play an important role. However, the migration effect and molecular process of decidual stromal cell on trophoblastic cells remains further investigation. In our study, we aimed to observe the effects of deciduas on the invasion and migration of trophoblastic cells and the impacts on the related gene (including MMPs and TIMPs,) expression. Methods:By primary cultured we established human decidual stromal cells and human endometrial mesenchymal cells. And immunofluorescencewas carried out to detect the expression of epithelial marker cytokeratin and mesenchymal marker vimentin these cells. Culture condition medium from decidual stromal cells and human endometrial mesenchymal cells were collected respectively, which were termed as DSC-CM and EMC-CM accordingly. Trophoblastic cancer cells JAR were pretreated for48hours by DSC-CM and EMC-CM respectively. Then cell migration abilities were evaluated by transwell assay, comparing the cells numbers migrated through the chamber membrance. Q-PCR and Western blot was applied to detect the expression level of MMPs and TIMPs in both groups.Results:Our immunofluorescence staining results showed that up to95%of the cultured cells were vimentin positive and cytokeratin negative. And lower than5%of them were vimentin negative and cytokeratin positive. These results identified that a majority of our primary cultured cells were mesenchymal cells, which promised the success of our cells establishment. By transwell assays, we found that cell numbers of DSC-CM pretreated trophoblastic cells migrated through chamber membrane were significantly greater than EMC-CM pretreated cells or blank cells, which indicated that more powerful migration inducible ability of decidual stromal cells compared with endometrial mesenchymal cells. To verify this phenomenon, we carried out Q-PCR and Western blot to determine the expression level of MMPs and TIMPs in condition medium pretreated trophoblastic cells. Our data showed that DSC-CM obviously increased JAR MMP2and MMP9expression level, while TIMP1and TIMP2mRNA have no significant difference between groups.Conclusion:Our results found that compared to EMC-CM, DSC-CM promoted the migration of trophoblastic cells, and raised the expression level of MMP2and MMP9, which indicated that DSC-CM enhance trophoblast migration ability probably by inducing MMP2and MMP9expression.