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Several chondrichthyan species are threatened, and we must increase our knowledge of their reproductive biology in order to establish assisted reproductive protocols for ex situ or in situ endangered species. The small-spotted catshark (<i>Scyliorhinus canicula</i>) is one of the most abundant shark species of the Mediterranean coast and is easy to maintain in aquaria; therefore, it is considered an ideal reproductive model. This study aimed to compare <i>S. canicula</i> male reproductive function in aquarium-housed (<i>n</i> = 7) and wild-captured animals, recently dead (<i>n</i> = 17). Aquarium-housed animals had lower semen volume (<i>p</i> = 0.005) and total sperm number (<i>p</i> = 0.006) than wild-captured animals, but similar sperm concentrations. In terms of sperm parameters, aquarium-housed sharks showed higher total sperm motility (<i>p</i> = 0.004), but no differences were observed regarding sperm viability, mitochondrial membrane potential, or membrane integrity. A morphometric study pointed to a significantly longer head (<i>p</i> = 0.005) and acrosome (<i>p</i> = 0.001) in wild-captured animals. The results of the spermatozoa morphological study of <i>S. canicula</i> were consistent with previous results obtained in other chondrichthyan species. With regard to sex hormones, testosterone levels were significantly lower in aquarium-housed animals (<i>p</i> ≤ 0.001), while similar levels of 17β-estradiol and progesterone were found. In short, the present study provides evidence of good in vitro semen quality in <i>S. canicula</i> housed in an aquarium, underlining their excellent potential for application in reproductive technologies for this and other chondrichthyan species.