Fachbereich Veterinärmedizin



    Establishment and characterization of a differentiated epithelial cell culture model derived from the porcine cervix uteri (2012)

    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Miessen, Katrin
    Einspanier, Ralf (WE 3)
    Schoen, Jennifer
    BMC veterinary research; 8(1) — S. 31
    ISSN: 1746-6148
    URL (Volltext): http://edocs.fu-berlin.de/docs/receive/FUDOCS_document_000000014772
    DOI: 10.1186/1746-6148-8-31
    Pubmed: 22429795
    Institut für Veterinär-Biochemie

    Oertzenweg 19 b
    14163 Berlin
    Tel.+49 30 838 62225 Fax.+49 30 838-62584

    Abstract / Zusammenfassung

    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.

    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.

    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.