Isolation and Characterization of Goat Endometrial Cells

Primary goat endometrial epithelial cells (EECs) and stromal cells (ESCs) were isolated from the endometrium of 8-month-old goats (n = 6) on Day 7 of estrous cycle as previously described [11 (link)]. The primary endometrial cells were cultured and maintained in DMEM/F-12 (Gibco, Invitrogen Corporation, Grand Island, NY) supplemented with 10% charcoal stripped fetal calf serum (FCS; Biowest, France), antibiotics (100 U/ml penicillin, 100 μg/ml streptomycin; Gibco) at 37°C in a 5% CO₂ –95% air atmosphere. The primary endometrial cells cultured on the cover slides were immediately fixed in 95% ethanol (v/v) and immunostained for cytokeratin and vimentin using an avidin-biotin complex (ABC) immunoperoxidase kit (Vectastatin Elite ABC kit; Vector Labs, Burlingame, CA) [11 (link)]. The protein expressions of 17β-estradiol receptor (ER) and progesterone receptor (PR) were analyzed separately in the EECs and ESCs using western blotting as previously described [12 (link)].