Global expansion in fish production and trade of aquatic ornamental species requires advances in aquatic animal health management. Aquatic parasite cultures permit diverse research opportunities to understand parasite–host dynamics and are essential to validate the efficacy of treatments that could reduce infections in captive populations. Monogeneans are important pathogenic parasites of captured captive fishes and exhibit a single-host life cycle, which makes them amenable to in vivo culture. Continuous cultures of oviparous monogenean parasites provide a valuable resource of eggs, oncomiracidia (larvae) and adult parasites for use in varied ecological and applied scientific research. For example, the parasite–host dynamics of Entobdella soleae (van Beneden and Hesse, 1864) and its fish host, Solea solea (Linnaeus, 1758), is one of the most well-documented of all monogeneans following meticulous, dedicated study. Polystoma spp. cultures provide an intriguing model for examining evolution in monogeneans because they exhibit two alternative phenotypes depending on the age of infection of amphibians. Furthermore, assessments of the ecological, pathological and immunological effects of fish parasites in aquaculture have been achieved through cultures of Gyrodactylus von Nordmann, 1832 spp., Benedenia seriolae (Yamaguti, 1934), Neobenedenia Yamaguti, 1963 spp. and Zeuxapta seriolae (Meserve, 1938). This review critically examines methods to establish and maintain in vivo monogenean monocultures on finfish, elasmobranchs and amphibians. Four separate approaches to establish cultures are scrutinised including the collection of live infected hosts, cohabiting recipient hosts with infected stock, cohabiting hosts with parasite eggs or oncomiracidia (larvae) and direct transfer of live adult parasites onto new fish hosts. Specific parasite species’ biology and behaviour permits predictive collection of parasite life stages to effectively maintain a continuous culture, while environmental parameters can be altered to manipulate parasite generation time. Parasite virulence and biosecurity are vital components of a well-managed culture to ensure appropriate animal welfare and uncontaminated surrounding environments. Contemporary approaches and techniques are reviewed to ensure optimised monogenean cultures, which ultimately can be used to further our understanding of aquatic parasitology and identify mechanisms to limit infestations in public aquaria, ornamental trade and intensive aquaculture.