Expression of adhesion and extracellular matrix genes in human blastocysts upon attachment in a 2D co-culture system

AbstractSTUDY QUESTIONWhat are the changes in human embryos, in terms of morphology and gene expression, upon attachment to endometrial epithelial cells?SUMMARY ANSWERApposition and adhesion of human blastocysts to endometrial epithelial cells are predominantly initiated at the embryonic pole and these steps are associated with changes in expression of adhesion and extracellular matrix (ECM) genes in the embryo.WHAT IS KNOWN ALREADYBoth human and murine embryos have been co-cultured with Ishikawa cells, although embryonic gene expression associated with attachment has not yet been investigated in an in vitro implantation model.STUDY DESIGN, SIZE, DURATIONVitrified human blastocysts were warmed and co-cultured for up to 48 h with Ishikawa cells, a model cell line for receptive endometrial epithelium.PARTICIPANTS/MATERIALS, SETTING, METHODSSix days post-fertilization (6dpf) human embryos were co-cultured with Ishikawa cells for 12, 24 (7dpf) or 48 h (8dpf) and attachment rate and morphological development investigated. Expression of 84 adhesion and ECM genes was analysed by quantitative PCR. Immunofluorescence microscopy was used to assess the expression of three informative genes at the protein level. Data are reported on 145 human embryos. Mann –WhitneyU was used for statistical analysis between two groups, withP< 0.05 considered significant.MAIN RESULTS AND THE ROLE OF CHANCEThe majority of embryos attached to Ishikawa cells at the level of the polar trophectoderm; 41% ...
Source: Molecular Human Reproduction - Category: Molecular Biology Source Type: research