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

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 alpha-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.

Polarised bovine endometrial epithelial cells vectorially secrete prostaglandins and chemotactic factors under physiological and pathological conditions

“…The stabilised TER identified in this study after w5 days of culture was comparable to a bovine caruncular epithelial cell line (Bridger et al 2007). The finding that polarised endometrial epithelial monocultures can establish high TER is in agreement with reports for mouse endometrial cells (Grant & Wira 2003, Grant-Tschudy & Wira 2005b.…”

“…Regulation of tight junctions is important for the function of epithelia under physiological and pathological conditions (Balda et al 1992, Matter et al 2005. A bovine caruncular cell line (BCEC-1) established confluence when TER was 600-1600 Ucm 2 , depending on the coating matrix of the insert (Bridger et al 2007), and the presence of stromal cells was essential for the formation of tight junctions in cultured bovine endometrial epithelial cells (Ulbrich et al 2011). However, the impact of epithelial cell polarisation on the mechanisms of bovine endometrial physiology or pathology is less well established.…”

Establishment and Characterization of Porcine Colonic Epithelial Cells Grown in Primary Culture

“…(4) DMEM/F12 mix (Sigma-Aldrich): like DMEM 4.5, the DMEM/F12 mix has also been used successfully to culture epithelial cells [Aldhous et al, 2001;Schierack et al, 2006;Bridger et al, 2007;Koh et al, 2008]. Thus, we tested whether it would be more suitable for culturing colonic epithelial cells than DMEM 4.5.…”

Epidermal growth factor (EGF) induces motility and upregulates MMP‐9 and TIMP‐1 in bovine trophoblast cells

“…Since the underlying mechanisms regulating these processes are barely understood, we recently established a trophoblast cell line (F3) from bovine placentomes. This tool allows the evaluation of possible differences and regulatory mechanisms in both compartments in vitro (Bridger et al, 2007;Hambruch et al, 2010).…”