title: Role for the endometrial epithelial protein MFG-E8 and its receptor integrin αvβ3 in human implantation: results of an in vitro trophoblast attachment study using established human cell lines.
abstract: OBJECTIVE
  To investigate the role of MFG-E8 and its receptor integrin αvβ3 in the attachment of trophoblast cells to the endometrial epithelium.
  
  
  METHODS
  Experimental in vitro study.
  
  
  METHODS
  Academic center.
  
  
  METHODS
  None.
  
  
  METHODS
  By using a well-differentiated endometrial adenocarcinoma cell line (Ishikawa cells) and choriocarcinoma human trophoblast cells (Jar cells), an in vitro assay mimicking human implantation was established. To investigate the impact of blocking MFG-E8 and integrin αvβ3, we pretreated the cell lines with antibodies against those proteins at different concentrations before the attachment assay.
  
  
  METHODS
  Attachment rate of Jar spheroids to the epithelial cell monolayer.
  
  
  RESULTS
  Pretreatment of Ishikawa cells with anti-MFG-E8 antibody caused a dose-dependent and significant inhibition of attachment. On the other hand, pretreatment of Jar spheroids did not result in a significant effect on the attachment rate. Pretreatment of Ishikawa cells as well as Jar spheroids with anti-integrin αvβ3 antibodies resulted in a dose-dependent, significant inhibition of attachment.
  
  
  CONCLUSIONS
  This study showed that blocking MFG-E8 and its receptor integrin αvβ3 in Ishikawa cells diminishes Jar spheroid attachment. Moreover, blocking integrin αvβ3 in the trophoblastic cells also diminished their attachment to the Ishikawa monolayer.
authors.affiliation: Department of Obstetrics and Gynecology, Eastern Virginia Medical School, The Jones Institute for Reproductive Medicine, Norfolk, Virginia.
authors.forename: Carla
authors.initials: C
authors.lastname: Schmitz
authors.affiliation: Department of Obstetrics and Gynecology, Eastern Virginia Medical School, The Jones Institute for Reproductive Medicine, Norfolk, Virginia.
authors.forename: Liang
authors.initials: L
authors.lastname: Yu
authors.affiliation: Department of Obstetrics and Gynecology, Eastern Virginia Medical School, The Jones Institute for Reproductive Medicine, Norfolk, Virginia.
authors.forename: Silvina
authors.initials: S
authors.lastname: Bocca
authors.affiliation: Department of Obstetrics and Gynecology, Eastern Virginia Medical School, The Jones Institute for Reproductive Medicine, Norfolk, Virginia.
authors.forename: Sandra
authors.initials: S
authors.lastname: Anderson
authors.affiliation: Department of Obstetrics and Gynecology, Eastern Virginia Medical School, The Jones Institute for Reproductive Medicine, Norfolk, Virginia.
authors.forename: João Sabino
authors.initials: JS
authors.lastname: Cunha-Filho
authors.affiliation: Department of Mathematics and Statistics, Old Dominion University, Norfolk, Virginia.
authors.forename: Bhaskara S
authors.initials: BS
authors.lastname: Rhavi
authors.affiliation: Department of Obstetrics and Gynecology, Eastern Virginia Medical School, The Jones Institute for Reproductive Medicine, Norfolk, Virginia. Electronic address: Oehninsc@evms.edu.
authors.forename: Sergio
authors.initials: S
authors.lastname: Oehninger
mesh_terms: D000912:Antibodies, Neoplasm; D000954:Antigens, Surface / Q000276:immunology / Q000502:physiology*; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D010064:Embryo Implantation / Q000276:immunology*; D004717:Endometrium / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D008894:Milk Proteins / Q000037:antagonists & inhibitors / Q000276:immunology; D011247:Pregnancy; D014327:Trophoblasts / Q000378:metabolism*
chemical_list: D000912:Antibodies, Neoplasm; D000954:Antigens, Surface; D039302:Integrin alphaVbeta3; C098942:MFGE8 protein, human; D008894:Milk Proteins
keywords: Attachment; Jar spheroids; MFG-E8; human endometrium; implantation; integrin αvβ3