Abstract: Cell culture technology has made important contributions since 19th century,in various disciplines such as genetics,immunology,experimental medicine,and oncology.Cell culture of aquatic animals began in the 1960s.It was originally used for the isolation and purification of viruses and is of great significance for the prevention and treatment of diseases in aquatic animals.Three-dimensional(3D) culture of cells is a new culture method developed based on traditional cell culture technology,which can better simulate the growth environment and condition of cells in living organisms,and has gained more and more attention.Based on the summary of the characteristics and development of 3D cell culture technology,this paper briefly introduced and prospected the application of 3D cell culture technology in aquaculture.