Isolation and culture of bovine endometrial epithelial cells in a serum-free culture system

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.