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Porcine Endometrial Glandular Epithelial Cells in Vitro: Transcriptional Activities of the Pregnancy-Associated Genes Encoding Antileukoproteinase and Uteroferrin1

1996; Oxford University Press; Volume: 55; Issue: 2 Linguagem: Inglês

10.1095/biolreprod55.2.469

1529-7268

Karen L. Reed, L. Badinga, Duane L Davis, Tricia E. Chung, Rosalia C.M. Simmen,

Reproductive Physiology in Livestock

The aim of this investigation was to establish a homologous culture system for study of the transcriptional mechanisms underlying endometrial expression of the pregnancy-associated genes encoding antileukoproteinase (ALP), an elastase/cathepsin G protease inhibitor, and uteroferrin (Uf), a transplacental iron transport protein. Glandular epithelial (GE), Luminal epithelial (LE), and stromal (ST) cells were isolated from pig endometrium at Day 12 of pregnancy by differential enzymatic digestion and sieve filtration. The three cell populations differed with respect to their morphology in culture and with respect to their expression of ALP and Uf. Expression of the ALP gene was much higher in GE than in LE cells and was undetectable in ST cells. Similarly, GE had the highest expression of the Uf gene, and expression in ST was lower but distinct. Western blot analysis of conditioned media (72 h) from GE, LE, and ST, using antiporcine Uf antiserum, detected significant levels of secreted Uf only in GE. The steroid hormone responsiveness of GE cells was monitored by changes in steady-state levels of ALP mRNA after 24-h exposure to estradiol 17β (E2; 10 nM) and/or progesterone (P; 10 nM). Glandular epithelial cells treated with E2, P, and E2 + P had increased (p < 0.05) ALP mRNA levels relative to those in control cultures.