Establishing primary cell cultures from the mesopelagic and deep-sea fishes is challenging, due to the adaptations of the organisms to high pressure, low temperature, and the logistical difficulties in retrieving specimens to surface. In the present study, we attempted to establish primary cell cultures from the fishes collected at a depth range from 100 to 250 m, by a single-day commercial trawler. Primary cultures were initiated through explant method, for live fishes (Sorsogona tuberculata, Platycephalus indicus, Uranoscopus marmoratus, and Halieutaea sp.) and the onboard-process of Priacanthus hamrur (caudal and pectoral fins), following sequential antibiotic washes. Among the 16 tissues attempted, cell outgrowth was observed in the heart of S. tuberculata, eye of P. indicus, eye and heart of U. marmoratus, and the caudal and pectoral fins of P. hamrur. Sustained proliferation beyond five passages was achieved only for U. marmoratus (eye and heart) and P. hamrur (caudal and pectoral fins). P. hamrur cells were maintained for over eight passages, and the findings demonstrate the feasibility of deriving primary cultures from deep-water fish tissues without specialized pressure-retention systems, offering a practical foundation for future studies on cellular and molecular adaptations of organisms inhabiting deep-water environments.