title: Establishment and characterization of a differentiated epithelial cell culture model derived from the porcine cervix uteri.
abstract: BACKGROUND
  Cervical uterine epithelial cells maintain a physiological and pathogen-free milieu in the female mammalian reproductive tract and are involved in sperm-epithelium interaction. Easily accessible, differentiated model systems of the cervical epithelium are not yet available to elucidate the underlying molecular mechanisms within these highly specialized cells. Therefore, the aim of the study was to establish a cell culture of the porcine cervical epithelium representing in vivo-like properties of the tissue.
  
  
  RESULTS
  We tested different isolation methods and culture conditions and validated purity of the cultured cells by immunohistochemistry against keratins. We could reproducibly culture pure epithelial cells from cervical tissue explants. Based on a morphology score and the WST-1 Proliferation Assay, we optimized the growth medium composition. Primary porcine cervical cells performed best in conditioned Ham's F-12, containing 10% FCS, EGF and insulin. After cultivation in an air-liquid interface for three weeks, the cells showed a discontinuously multilayered phenotype. Finally, differentiation was validated via immunohistochemistry against beta catenin. Mucopolysaccharide production could be shown via alcian blue staining.
  
  
  CONCLUSIONS
  We provide the first suitable protocol to establish a differentiated porcine epithelial model of the cervix uteri, based on easily accessible cells using slaughterhouse material.
authors.affiliation: Freie Universität Berlin, Institute of Veterinary Biochemistry, Oertzenweg 19b, Berlin, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Miessen
authors.forename: Ralf
authors.initials: R
authors.lastname: Einspanier
authors.forename: Jennifer
authors.initials: J
authors.lastname: Schoen
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000166:cytology*; D003470:Culture Media; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D006025:Glycosaminoglycans / Q000378:metabolism; D007633:Keratins / Q000378:metabolism; D013552:Swine; D051176:beta Catenin / Q000378:metabolism
chemical_list: D003470:Culture Media; D006025:Glycosaminoglycans; D051176:beta Catenin; D007633:Keratins