ISOLATION, CULTURE, AND CHARACTERIZATION OF EQUINE OVIDUCT EPITHELIAL-CELLS IN-VITRO

Oviduct epithelial cells (OEC) increasingly are used to support embryo nic development and to study gamete interactions with the female repro ductive tract in vitro. This series of experiments was designed to cha racterize monolayers derived from oviduct epithelium. Epithelial cells harvested from the isthmus and ampulla of the oviducts of five estrou s mares were cultured with or without the basal lamina extract, Matrig el. Within each group OEC were cultured in the presence of either estr adiol-17 beta or a carrier control. All groups were subcultured three times. Epithelial cell morphology and function were examined by micros copy, analysis of secreted proteins, and immunocytochemistry. Epitheli al cells attached more rapidly and reached confluence sooner when cult ured on Matrigel than in uncoated wells. Cells showed variable evidenc e of ciliary activity up to 12 days in primary culture. Cells grown on Matrigel had a more polarized appearance in primary culture than thos e in uncoated wells, although no morphologic difference between anatom ic site of origin or between steroid treated groups was noted. Anatomi c site of origin had no effect, and steroid treatment had minimal effe cts, on patterns of secreted proteins. However, some differences were noted in protein secretion between cells grown with or without Matrige l. These data suggest that culture substrate may affect structure and function of OEC monolayers. (C) 1995 Wiley-Liss, Inc.