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Establishment and characterization of a differentiated epithelial cell culture model derived from the porcine cervix uteri
Miessen, Katrin ;  Einspanier, Ralf ;  Schoen, Jennifer

HaupttitelEstablishment and characterization of a differentiated epithelial cell culture model derived from the porcine cervix uteri
AutorMiessen, Katrin
AutorEinspanier, Ralf
AutorSchoen, Jennifer
DDC630 Landwirtschaft, Veterinärmedizin
Auch erschienen inBMC Veterinary Research 2012, 8:31
ZusammenfassungBackground
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.
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Fachbereich/EinrichtungFB Veterinärmedizin
Arbeitsbereich/InstitutInstitute of Veterinary Biochemistry
Erscheinungsjahr2012
Dokumententyp/-SammlungenWissenschaftlicher Artikel
SpracheEnglisch
Rechte Creative Commons License
Dieses Werk ist unter einer Creative Commons-Lizenz lizenziert.
Anmerkungen des AutorsGefördert durch die DFG und den Open Access Publikationsfonds der Freien Universität Berlin
Erstellt am30.10.2012 - 09:02:50
Letzte Änderung10.01.2014 - 10:24:24
 
Statische URLhttp://edocs.fu-berlin.de/docs/receive/FUDOCS_document_000000014772
DOI10.1186/1746-6148-8-31
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