Bovine caruncular epithelial cell line (BCEC-1) isolated from the placenta forms a functional epithelial barrier in a polarised cell culture model

被引:23
作者
Bridger, P. S.
Menge, C.
Leiser, R.
Tinneberg, H. -R.
Pfarrer, C. D.
机构
[1] Univ Giessen, Dept Obstet & Gynecol, D-35385 Giessen, Germany
[2] Univ Giessen, Dept Vet Anat Histol & Embryol, D-35392 Giessen, Germany
[3] Univ Giessen, Inst Hyg & Infect Dis Anim, D-35392 Giessen, Germany
关键词
bovine placenta; cell culture; polarised epithelial cell line; transepithelial electrical resistance; bovine virus diarrhoea virus;
D O I
10.1016/j.placenta.2007.07.002
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 a-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 (R) 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. (c) 2007 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1110 / 1117
页数:8
相关论文
共 46 条
[1]   Experimental infection of pregnant cows with Bacillus licheniformis bacteria [J].
Agerholm, JS ;
Jensen, NE ;
Dantzer, V ;
Jensen, HE ;
Aarestrup, FM .
VETERINARY PATHOLOGY, 1999, 36 (03) :191-201
[2]  
ARSLAN A, 1995, IN VITRO CELL DEV-AN, V31, P140
[3]   In vitro response to oxytocin and interferon-tau in bovine endometrial cells from caruncular and inter-caruncular areas [J].
Asselin, E ;
Drolet, P ;
Fortier, MA .
BIOLOGY OF REPRODUCTION, 1998, 59 (02) :241-247
[4]   Expression of prostaglandin transporter in the bovine uterus and fetal membranes during pregnancy [J].
Banu, SK ;
Arosh, JA ;
Chapdelaine, P ;
Fortier, MA .
BIOLOGY OF REPRODUCTION, 2005, 73 (02) :230-236
[5]   Bovine interferon-τ stimulates the Janus kinase-signal transducer and activator of transcription pathway in bovine endometrial epithelial cells [J].
Binelli, M ;
Subramaniam, P ;
Diaz, T ;
Johnson, GA ;
Hansen, TR ;
Badinga, L ;
Thatcher, WW .
BIOLOGY OF REPRODUCTION, 2001, 64 (02) :654-665
[6]   Interferon-τ modulates phorbol ester-induced production of prostaglandin and expression of cyclooxygenase-2 and phospholipase-A2 from bovine endometrial cells [J].
Binelli, M ;
Guzeloglu, A ;
Badinga, L ;
Arnold, DR ;
Sirois, J ;
Hansen, TR ;
Thatcher, WW .
BIOLOGY OF REPRODUCTION, 2000, 63 (02) :417-424
[7]  
BRIDGER PS, 2007, IN PRESS THERIOGENOL
[8]   Platelet-activating factor receptor (PAF-R) and acetylhydrolase (PAF-AH) are co-expressed in immature bovine trophoblast giant cells throughout gestation, but not at parturition [J].
Bücher, K ;
Leiser, R ;
Tiemann, U ;
Pfarrer, C .
PROSTAGLANDINS & OTHER LIPID MEDIATORS, 2006, 79 (1-2) :74-83
[9]   Assessment of transport rates of proteins and peptides across primary human alveolar epithelial cell monolayers [J].
Bur, Michael ;
Huwer, Hanno ;
Lehr, Claus-Michael ;
Hagen, Nina ;
Guldbrandt, Mette ;
Kim, Kwang-Jin ;
Ehrhardt, Carsten .
EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES, 2006, 28 (03) :196-203
[10]  
ClassenLinke I, 1997, CELL TISSUE RES, V287, P171