The epithelium of the human epididymis maintains an appropriate luminal environment for sperm maturation that is essential for male fertility.Regional expression of small noncoding RNAs such as microRNAs contributes to segment-specific gene expression and differentiated functions.MicroRNA profiles were reported in human epididymal tissues but not specifically in the epithelial cells derived from those regions.Here,we reveal miRNA signatures of primary cultures of caput,corpus,and cauda epididymis epithelial cells and of the tissues from which they were derived.We identify 324 epithelial cell-derived microRNAs and 259 tissue-derived microRNAs in the epididymis,some of which displayed regionalized expression patterns in cells and/or tissues.Caput cell-enriched miRNAs included miR-573 and miR-155.Cauda cell-enriched miRNAs included miR-1204 and miR-770.Next,we determined the gene ontology pathways associated with in silico predicted target genes of the differentially expressed miRNAs.The effect of androgen receptor stimulation on miRNA expression was also investigated.These data show novel epithelial cell-derived miRNAs that may regulate the expression of important gene networks that are responsible for the regionalized gene expression and function of the epididymis.