In vitro systems for intercepting early embryo-maternal cross-talk in the bovine oviduct

Abstract

A comprehensive understanding of the complex embryo-maternal interactions during the preimplantation period requires the analysis of very early stages of pregnancy. These are difficult to assess in vivo due to the small size of the embryo exerting local paracrine effects. Specifically designed experiments and holistic transcriptome and proteome analyses to address the early embryo-maternal cross-talk in the oviduct require sufficient numbers of well-defined cells in a standardized experimental environment. The pronounced estrous cycle-dependent changes in gene expression and morphology of bovine oviduct epithelial cells (BOECs) clearly show that a precise definition of the stage of estrous cycle is essential for obtaining a well-defined homogenous population of functional cells. The number of intact cells isolated from individual ampullae by solely mechanical means was 10-fold higher than previously reported cell yields after enzymatic treatment, and the purity was comparable. Bovine oviduct epithelial cells have been cultured as monolayers or in suspension. Proliferating cells grown in monolayers dedifferentiated, with a concomitant loss of important morphologic characteristics. After several days in culture, BOECs in monolayers are less likely to mimic the oviduct environment in vivo than BOEC vesicles formed of epithelial sheets in short-term suspension culture. A 24-h culture system for BOECs isolated on Day 3.5 of the estrous cycle showed excellent preservation of morphologic criteria, marker gene expression, and hormone responsiveness. The short-term BOEC culture system provides well-defined and functional BOECs in sufficient quantities for studies of early embryo-maternal interactions in experiments that mimic the environment in the oviduct in vivo.