Publikationsdatenbank

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

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Miessen, Katrin

Einspanier, Ralf (WE 3)

Schoen, Jennifer

Quelle

BMC veterinary research

Bandzählung: 8

Heftzählung: 1

Seiten: 31

ISSN: 1746-6148

Sprache

Englisch

Verweise

URL (Volltext): http://edocs.fu-berlin.de/docs/receive/FUDOCS_document_000000014772

DOI: 10.1186/1746-6148-8-31

Pubmed: 22429795

Kontakt

Institut für Veterinär-Biochemie

Oertzenweg 19 b
14163 Berlin
+49 30 838 62225
biochemie@vetmed.fu-berlin.de

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.