%0 Journal Article
%T Establishment and characterization of a differentiated epithelial cell culture model derived from the porcine cervix uteri
%A Katrin Miessen
%A Ralf Einspanier
%A Jennifer Schoen
%J BMC Veterinary Research
%D 2012
%I BioMed Central
%R 10.1186/1746-6148-8-31
%X 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.The uterine cervical epithelium protects the upper reproductive tract from insults providing a physical barrier, secretions containing bactericidal and virucidal agents and a pathogen-dependent direct immunomodulation [1-3]. During estrous, it takes part in direct sperm-epithelium interaction [4] as well as in the signal reception from seminal fluid [5].To elucidate cell type-specific actions of hormones and cytokines, signal transduction pathways, cell-cell interactions, and gene expression in these highly specialized cells, model systems resembling the original tissue need to be developed.Cervical cell cultures of a variety of species are already applied in various fields of science. They serve as in vitro systems for basic research [6], in oncological and microbiological studies [7-9] as well as for assessment of product-and pharmaco-toxicity [10,11].The cells used in these studies are mainly derived from human ectocervical tissue, which in vivo is covered by a polarized, multilayered epithelium. However, the cells (primary, immortalized or transformed) are cultured as monolayers
%U http://www.biomedcentral.com/1746-6148/8/31