Immunohistochemical analysis of steroid receptors and glycodelin A (PP14) in isolated glandular epithelial cells of normal human endometrium

Highly purified fractions of isolated endometrial cells can be useful for i nvestigating endometrial function. After a first collagenase digestion, nor mal human endometrial stromal and epithelial cells were separated by filtra tion. Glands were purified further by two collagenase digestion steps, filt ration, differential sedimentations, and Ficoll gradient centrifugation. Ep ithelial cells were polyhedral and grew as islands in a whorl-like wavy pat tern around glandular fragments. High cell culture purity was confirmed wit h the positive immunohistochemical reaction against cytokeratin 7,8,18,19. Isolated human glands had a similar distribution pattern of estrogen recept or (ER) and progesterone receptor (PR) as observed in vivo, suggesting that glands have a functional hormone receptor system at the time of plating. U sing a specific monoclonal antibody against glycodelin A (GdA), a character istic cyclical expression was demonstrated during the menstrual cycle. The GdA reaction was weak in the proliferative phase, increasing significantly till the late secretory phase, suggesting a similar GdA concentration in vi tro as observed in vivo glands. In conclusion, this method could be a model for studying endometrial glandular cells from different menstrual phases, endometrial cell interactions, implantation mechanisms, GdA regulation mech anisms, and pharmacological or other influences on ER and PR alteration.