Abstract: Objective Culture, expansion and characterization of pseudomyxoma peritonei cells, which laid the foundation for the study of pseudomyxoma peritonei and drug screening. Methods Tumors from 5 cases of human pseudomyxoma peritonei were cultured by collagenase digestion. The cultured cells were then identified as tumor cells by chromosome karyotyping. The secretion of neutral mucopolysaccharide was detected by PAS staining. To simulate the tumor growth condition in vivo, 3D culture was applied and the morphology of the cultured organoid was observed with HE staining, xenografts were used to test the tumorigenicity of PMP primary cells. Results The primary culture of pseudomyxoma peritonei cells was successful in all of the 5 cases. 2D cultured cells were adherent, polygonal pebble-like arranged, and passed on up to 18 generations. Chromosome karyotype analysis showed that most of the cells were subdiploid karyotype tumor cells. Positive PAS staining suggests that these cells secrete mucus. The morphology of 3D cultured organoid was similar to that of tumor tissues. The tumor formation rate of xenografts was low, and the tumoris not similar to patient’s PMP. Conclusion Pseudomyxoma peritonei tumor cells can be cultured and expanded in vitro.

Key words: Key words：Pseudomyxoma peritonei, Primary culture , 3D culture