The invention discloses a culture method of oral squamous
cell carcinoma organoid. The culture method comprises the following steps: S1, preparing a dissociation
enzyme; S2, preparing a culture medium, wherein the culture medium is composed of a basal culture medium DMEM / F12, an R-spondin1 conditional culture medium, a Wnt3A conditional culture medium,
sterile water and functional components; S3, obtaining a sample through clinical operation, namely obtaining a tumor specimen through material taking or
surgical resection,
cutting tissue, washing the tissue with sterile normal
saline for three times, soaking the tissue in a DMEM medium for 1 hour and continuing to dissociate; S4, tissue pretreatment, namely placing the sample obtained in the step S3 in a sterile 6-well plate, and
cutting up sample tissue blocks by using a disposable sterile
surgical blade; S5, tissue dissociation and
digestion, namely adding the dissociation
enzyme prepared in the step S1, repeating the process of
centrifugation-supernatant removal for three times so as to fully remove the dissociation
enzyme, and mixing the cells with the sample tissue blocks before the last time of
centrifugation; and S6, carrying out
organoid culture.