Abstract: In vitro cell culture is an important experimental method to explore the occurrence and development of acute leukemia (AL).However,the traditional two-dimensional culture can not demonstrate the complex bone marrow structure.The three-dimensional cell culture model allows leukemia cells to grow in all directions in vitro,and to form a suitable spatial environment and cell connection between cells,which can effectively simulate the microenvironment of leukemia in vivo,and provide optimal condition for studying the biological characteristics of leukemia cells.In recent years,more and more studies have applied three-dimensional culture models for drug screening in order to provide more effective potential therapeutic measures.This article summarizes the application of three-dimensional cell culture in the studies of AL in vitro in recent years.