Culture method for screening tumor organoids based on P53 mutation

Abstract

The invention relates to the technical field of organoid culture, and particularly discloses a culture method for screening tumor organoids based on P53 mutation. The culture method comprises the following steps: S1, unfreezing untreated tumor organoid to generate a cell suspension, adding the cell suspension into a basic culture medium, uniformly conducting mixing and centrifuging, and removing supernate to obtain a tumor organoid mixture; S2, adding matrigel to resuspend the tumor organoid mixture, and after resuspending, inoculating the organoid mixture into an organoid culture medium containing y27632 for culturing; and S3, after the liquid of the tumor organoid mixture is changed for the first time, absorbing and removing the organoid culture medium, and then adding a culture medium containing Nutlin3A for continuous culture. The culture method is low in cost and price, sequencing is not needed, the detection cost is saved in the early stage of culture of the tumor organoids, and accurate and simplified culture of the tumor organoids is achieved.

Description

technical field [0001] The invention relates to the technical field of organoid culture, in particular to a method for culturing tumor organoids based on P53 mutation screening. Background technique [0002] At present, with the continuous deepening of research on targeted drugs, a variety of drugs targeting specific oncogenes have entered the clinic, bringing good news to the majority of patients. However, since the essence of cancer is a disease affected by multiple gene mutations, patient tumors will continue to evolve as the disease progresses, resulting in tumor heterogeneity between patients and heterogeneity within the same tumor. Cancer heterogeneity adds uncertainty to targeted precision therapy, and is also one of the important reasons leading to resistance to cancer drug therapy. Therefore, it is of great practical significance to expand precision medicine for cancer heterogeneity, and the rise of tumor organoid technology provides a new and more effective applic...

AI Technical Summary

Problems solved by technology

Cells with p53 deficiency do not have this control and continue to divide even under unfavorable conditions

[0006] The main basis for selecting targeted drugs in clinical research and treatment is tumor gene sequencing. Although tumor organoid culture methods can accurately simulate the tumor microenvironment in vivo and obtain organoid models after stable passage, they cannot detect P53 gene mutations in advance. Therefore, genetic sequencing is needed to determine whether there is a P53 mutation in organoids

However, gene sequencing is expensive, complicated to operate, and requires a large sample size

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