Oral squamous cell carcinoma organoid culture medium and culture method

Abstract

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.

Description

technical field

[0001] The invention relates to the technical field of biomedicine, and more specifically relates to a culture medium and a culture method for oral squamous cell carcinoma organoids. Background technique

[0002] Oral squamous cell carcinoma (Oral Squamous Cell Carcinoma, OSCC) is the most common malignant tumor of the oral and maxillofacial region. In 2020, there will be about 377,700 new cases of OSCC in the world, and 177,800 deaths. The 5-year survival rate is only about 60%. For middle-advanced OSCC patients, especially recurrent / metastatic OSCC patients, chemotherapy-assisted comprehensive therapy is a standardized treatment plan. For example, cisplatin and paclitaxel are the first-line chemotherapy drugs, but the therapeutic effect is not satisfactory, and targeted therapy drugs are of great benefit to the patients. The treatment effect of OSCC also needs to be further verified. Therefore, there is an urgent need to develop a more effective individuali...

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