title: Differential effects of oestradiol and progesterone on proliferation and morphology of cultured bovine uterine epithelial and stromal cells.
abstract: The effect of oestrogen and progesterone on the proliferation of cultured bovine uterine epithelial and stromal cells was assessed. Epithelial and stromal cells recovered from cows at day 1 to day 3 of the oestrous cycle were cultured in RPMI medium supplemented with 5% steroid-free fetal calf serum for 4 and 8 days. The addition of progesterone to the culture medium altered the morphology of stromal cells. Oestradiol (0.1-10 nmol l-1) and progesterone (50 nmol l-1) significantly increased the total DNA (from 9.6 +/- 0.96 to 25.6 +/- 0.99 micrograms per well, P < 0.001) and protein content (from 76.6 +/- 2.6 to 125.8 +/- 2.6 micrograms per well, P < 0.001) and decreased the ratio of protein to DNA (from 8.0 +/- 0.24 to 4.9 +/- 0.24, P < 0.01) in stromal cells during the first 4 days. During the second 4 days, the relative percentages of increase in DNA content were not affected by steroids, indicating that the major effect of steroids on stromal cell proliferation was exerted during the first 4 days of incubation. The morphology of epithelial cells was not influenced by the addition of steroids. DNA content of epithelial cells was reduced by the addition of oestrogen (from 22.9 +/- 2.1 to 15.0 +/- 2.0 micrograms per well, P < 0.01), but not progesterone (from 22.9 +/- 2.1 to 25.8 +/- 2.0 micrograms per well, P > 0.05). Total protein content of epithelial cells was reduced by oestradiol by day 4 (from 111.0 +/- 6.2 to 71.0 +/- 6.2 micrograms per well, P < 0.01), but not by day 8 (from 305.0 +/- 10.5 to 296.0 +/- 10.5 micrograms per well, P > 0.05). Progesterone increased the total protein content (from 305.0 +/- 10.5 to 366.0 +/- 10.5 micrograms per well, P < 0.01). Oestradiol significantly enhanced the ratio of protein to DNA in epithelial cells at day 8 (from 10.1 +/- 1.0 to 16.8 +/- 1.0, P < 0.01). These results show that oestradiol and progesterone have different effects on the proliferation and morphology of epithelial and stromal cells in vitro.
authors.affiliation: Centre de Recherche en Reproduction Animale, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Quebec, Canada.
authors.forename: C W
authors.initials: CW
authors.lastname: Xiao
authors.forename: A K
authors.initials: AK
authors.lastname: Goff
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology / Q000187:drug effects; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D005353:Fibronectins / Q000032:analysis; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis; D011374:Progesterone / Q000494:pharmacology*; D011506:Proteins / Q000032:analysis; D017154:Stromal Cells / Q000737:chemistry / Q000166:cytology / Q000187:drug effects; D013997:Time Factors; D014599:Uterus / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*
chemical_list: D005353:Fibronectins; D011506:Proteins; D011374:Progesterone; D004958:Estradiol; D007633:Keratins; D004247:DNA