Abstract: Establishment of appropriate tumor models has been an urgent need in cancer research. Established tumor models, including patient-derived tumor cell lines (PDC) and patient-derived tumor xenografts (PDX), have been widely used and facilitated tumor mechanism research and clinical application. But the loss of tumor heterogeneity in PDC limits its applications. Though PDX largely preserves tumor heterogeneity, the long culture period and low tumorigenesis rate make it costly. Due to the limitation above, the 3D culture system including multicellular tumor spheroids (MCTS) and patient-derived tumor organoids(PDO) has been developed. It largely mimics the interaction between tumor cells as well as the interaction between tumors and tumor environment, preserving tumor heterogeneity to a certain extent. Compared to PDX, MCTS and PDO are easier to establish and develop as a long-term passage or even a biobank, facilitating high-throughput drug screening and showing great potential in cancer research.