Bovine Caruncular Epithelial Cell Line (BCEC-1) Isolated from the Placenta Forms a Functional Epithelial Barrier in a Polarised Cell Culture Model

Bridger, P.S.; Menge, C.; Leiser, R.; Tinneberg, H.-R.; Pfarrer, C.D.

doi:10.1016/j.placenta.2007.07.002

« Previous Next »Placenta
Volume 28, Issue 11 , Pages 1110-1117, November 2007

Bovine Caruncular Epithelial Cell Line (BCEC-1) Isolated from the Placenta Forms a Functional Epithelial Barrier in a Polarised Cell Culture Model

P.S. Bridger
- Department of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, Frankfurter Strasse 98, D-35392 Giessen, Germany
C. Menge
- Institute for Hygiene and Infectious Diseases of Animals, Justus-Liebig-University, Frankfurter Strasse 85-89, D-35392 Giessen, Germany
R. Leiser
- Department of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, Frankfurter Strasse 98, D-35392 Giessen, Germany
H.-R. Tinneberg
- Department of Obstetrics and Gynecology, Justus-Liebig-University, Klinikstrasse 32, D-35385 Giessen, Germany
C.D. Pfarrer
- Department of Obstetrics and Gynecology, Justus-Liebig-University, Klinikstrasse 32, D-35385 Giessen, Germany
- Corresponding author. Tel.: +49 641 99 45282; fax: +49 641 99 45258.

Accepted 5 July 2007. published online 12 September 2007.

Abstract

In the bovine synepitheliochorial placenta key sites of fetal–maternal interaction are placentomes consisting of maternal caruncles interdigitating with fetal cotyledons. The aim of this study was to establish an epithelial cell line from caruncles of pregnant cows and to develop a model to study restricted trophoblast invasion, pathogenesis of pregnancy associated diseases and pathways of infection and transport. Primary epithelial cells were isolated, successfully subcultured for 32 passages and cryopreserved at various stages. The cultures were termed bovine caruncular epithelial cell line-1 (BCEC-1). Cytokeratin, zonula occludens-1 protein and vimentin but neither α-smooth muscle actin nor desmin were detected by immunofluorescence performed every 5 (±1) passages. These results were confirmed by Western blotting. BCEC-1 were then cultured either without matrix or on fibronectin or collagen coated Transwell^® polyester membrane inserts, respectively, enabling separate access to the basal or apical epithelial compartments. Transmission and scanning electron microscopy of BCEC-1 revealed ultrastructural features also observed in vivo, such as apical microvilli and junctional complexes. Transepithelial electrical resistance (TEER) was measured regularly and revealed an increase with advancing confluence in all cultures. Cultures on coated inserts reached confluence and corresponding TEER-levels at an earlier stage. In addition, the cells were tested negative for bovine virus diarrhoea (BVD) virus, but were permissive for the virus. In conclusion, the BCEC-1 cell line retained characteristics of maternal caruncular epithelial cells as observed in vivo and in primary cell cultures and thus will be a highly useful tool for future studies of pathways of invasion, fetal–maternal communication, transport and infection.

Keywords: Bovine placenta, Cell culture, Polarised epithelial cell line, Transepithelial electrical resistance, Bovine virus diarrhoea virus