Isolation and Culture of Bovine Endometrial Epithelial Cells in a Serum-Free Culture System.

Abstract

Abstract. The objective of this study was to establish a serum-free culture system for bovine endometrial epithelial cells, and to identify the function of these cells. Epithelial cells were separated from uterine endometrium at Days 5-10 of the estrous cycle by 0.76% EDTA-PBS and collagenase treatment. After histological observation, the epithelium was completely separated from the basement membrane by EDTA treatment. The separated epithelial cells were then dispersed by collagenase and cultured in Dulbecco's Modified Eagle's Medium/Ham's F-12 1:1 supplemented with insulin, transferrin, sodium-selenite, hydrocortisone, retinol, ascorbic acid and antibiotics on collagencoated culture dishes. The cells attached to the substrate and reached confluency after 5-7 days of culture. An immunocytochemical study revealed that the cultured epithelial cells were positive to cytokeratin, but not vimentin. The production of prostaglandin (PG) F2α was significantly higher (p<0.001) in the presence of 100 nM oxytocin (OT; 2.81 fold greater than the concentration in the control) than in the absence of OT. These results indicate that it is possible to culture bovine endometrial epithelial cells in the serum-free culture system, and that cultured cells have some typical characteristics of endometrial epithelial cells.

Biological Activity of Recombinant Bovine Interferon τ Produced by a Silkworm-Baculovirus Gene Expression System

“…In the present study, PGF 2α secretion by cultured epithelial cells was stimulated by OT, and the increase (to a level 2.45 times that in the control) was as great as reported in previous studies [32, 33]. Bm-rbIFNτ was shown to suppress the secretion of PGF 2α from cultured epithelial cells in a dose-dependent manner, and all concentrations (1 to 1,000 n g/m l ) of Bm-rbIFNτ significantly ( P <0.05) suppressed OT-induced secretion of PGF 2α (Fig.…”

A Passage and Storage System for Isolated Bovine Endometrial Epithelial and Stromal Cells

“…The production of PGF 2α in response to OT is a useful parameter for investigating the physiology and endocrine status of the cultured bovine endometrial epithelial cells [6,8,13]. In the present study, OT strongly stimulated PGF2α production in both, frozen and unfrozen primary cultures of the epithelial cells (Fig.…”

“…Several studies have described the isolation and primary culture of bovine endometrial cells [5][6][7][8]. However, Lopez-Gatius et al [9] reported a higher incidence of ovarian cysts during the warm period, which is in agreement with several investigations showing various disturbances of the estrous cycle and pregnancy in certain geographical areas [10][11][12].…”

Development of an in vitro Model for Bovine Placentation: A Comparison of the in vivo and in vitro Expression of Integrins and Components of Extracellular Matrix in Bovine Placental Cells

“…A further purification of epithelial cells was achieved by controlled incubation with trypsin/ EDTA (0.05/0.02%; w/v; Biochrom). Overgrowth of the epithelial cells by fibroblast cells due to FCS addition [Takahashi et al, 2001a] was not observed, because fibroblasts were present in very small numbers. Moreover, each passage further enriched epithelial cells by selective detachment of cells, i.e.…”