Development, optimization and use of a cell système from murine epididymal epithelium : molecular approaches
In mammals, the epididymal epithelium, through its activities and secretion reabsorption, participates in post-testicular maturation of sperm by creating a luminal environment particularly along the epididymal course. Studies to characterize the activities of secretion of this organ and to analyze gene regulation involved, have been hampered by the absence of cells in culture to this very epithelium differentiated. Despite numerous attempts, the systems of cultured cells developed so far do not reflect the state of cell differentiation epididymis in vivo. In particular, genes, which can be considered as terminal differentiation markers of epididymal, not expressed in cells in culture. GPX5, laboratory model gene, encoding a secreted protein of the family of glutathione peroxidases is a good example of epididymis-specific gene, that are not found in cells in culture systems available today. In this study, we used GPX5, PEA3 and indoleamine 2, 3-dioxygenase (IDO) and other genes expressed in the epididymis of mammals, as indicators of the state of differentiation of established cell lines head epididymis murine. These are derived from primary cultures, which are naturally immortalized by accumulation of random mutations. In a second step, we have tried to optimize the culture conditions of these cells to improve the level of epididymal gene expression, particularly GPX5, taken as reference.