After release from the testis (process known as spermiation), spermatozoa transit through the epididymis, a long-convoluted tubule that connects the efferent ducts to the vas deferens. During the transit, spermatozoa undergo several molecular modifications which are necessary to acquire progressive motility, to complete maturation and compaction of chromatin, as well as to regulate their volume and gain molecules necessary for fertilization. These modifications occur as spermatozoa pass through the three segments of epididymis, caput, corpus, and cauda, and are accomplished by secretions from epididymal epithelium. Among these secretions, epididymosomes (extracellular microvesicles enriched in cholesterol and proteins) and miRNA play important roles. Epididymal secretions are regulated by a variety of factors, primarily androgens and estrogens, which together create a distinct luminal environment in each epididymal segment. This environment supports progressive sperm maturation and ensures the maintenance of sperm viability and motility during storage in the cauda. Epididymal pH and electrolytes composition of the luminal fluid are also crucial for a correct sperm maturation. Finally, epididymal contractions allow sperm emission during ejaculation. Overall, the role of epididymis is essential for male reproduction, and alterations in any of its functions may lead to subfertility or infertility. Due to its importance for a successful male reproductive function, the epididymis appears to be a promising target for post-testicular male contraception.