title: Effect of primary human endometrial stromal cells on epithelial cell receptivity and protein expression is dependent on menstrual cycle stage.
abstract: BACKGROUND
  Successful implantation requires a receptive endometrium. We hypothesized that effects of endometrial stromal cells (ESC) on epithelial cell receptivity and trophoblast-endometrium interaction are menstrual cycle dependent.
  
  
  METHODS
  An endometrial in vitro 3D co-culture model of primary human ESC with the endometrial epithelial cell line (RL95-2) was constructed. Co-cultures were prepared using primary ESC from biopsies taken before the window of implantation (ESCbw) and during the window of implantation (ESCw), on cycle days 10-17 and 19-23, respectively. RL95-2 served as a constant parameter upon which the influence of ESC from different phases of the cycle was investigated. proMMP-2 (MMP, matrix metalloproteinase) and proMMP-9 secretion was tested in response to progesterone. Progesterone receptor B (PR-B) and plexin B1 protein expression and mRNA levels were investigated using immunofluorescence and RT-PCR, respectively.
  
  
  RESULTS
  Progesterone increased proMMP-2 secretion in primary ESCbw (P = 0.0046) but decreased proMMP-2 and proMMP-9 secretion in ESCw (P < 0.0005). In the presence of ESCbw, JAR spheroid attachment rate to overlying RL95-2 cells was decreased (P < 0.0001), whereas in the presence of ESCw, attachment rate was unchanged. Progesterone treatment restored epithelial cell receptivity in co-culture with ESCbw (P = 0.00004). A correlation between spheroid attachment rate and plexin B1 mRNA level was observed (P = 0.01). PR-B protein and mRNA level were influenced by the interplay between RL95-2 and stromal cells.
  
  
  CONCLUSIONS
  The effects of human primary ESC on epithelial cell receptivity and trophoblast-endometrium interaction depended upon whether the ESC were taken before or during the window of implantation.
authors.affiliation: Department of Obstetrics and Gynecology, Ha'Emek Medical Center, Afula, Israel.
authors.forename: A
authors.initials: A
authors.lastname: Evron
authors.forename: S
authors.initials: S
authors.lastname: Goldman
authors.forename: E
authors.initials: E
authors.lastname: Shalev
mesh_terms: D000328:Adult; D002450:Cell Communication; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D004717:Endometrium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004792:Enzyme Precursors / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D018093:Gelatinases / Q000378:metabolism; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008597:Menstrual Cycle / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D011374:Progesterone / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D011980:Receptors, Progesterone / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018874:Spheroids, Cellular; D017154:Stromal Cells / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism
chemical_list: D004792:Enzyme Precursors; D009419:Nerve Tissue Proteins; C492119:PLXNB1 protein, human; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D011980:Receptors, Progesterone; C097314:progesterone receptor B; D011374:Progesterone; D018093:Gelatinases; C120487:pro-matrix metalloproteinase 9; C069860:progelatinase; D020780:Matrix Metalloproteinase 9