Method for culturing bladder cancer organs in vitro

Abstract

The invention discloses a method for culturing bladder cancer organs in vitro. The method comprises the following steps: manufacturing a gas-liquid interaction culture system, namely manufacturing a uniformly paved rat tail collagen supporting layer on the surface of a porous culture membrane in a Transwell upper chamber; re-suspending fresh in-vitro bladder cancer tissues by using a rat tail collagen solution, uniformly mixing, adding the obtained mixture onto the rat tail collagen supporting layer, and then putting the rat tail collagen supporting layer into an incubator at 37 DEG C to solidify so as to obtain a rat tail collagen layer containing the bladder cancer tissues; adding an organoid culture medium into a Transwell lower chamber, and enabling the liquid level of the culture medium to be lower than the rat tail collagen layer containing bladder cancer tissues; and carrying out passage and cryopreservation. The culture success rate of the bladder cancer organoid is remarkablyincreased, the bladder cancer organoid with reserved immune cells can be obtained through culture, operation is easy, the utilization rate of tumor tissue is high, and great significance and value areachieved for bladder cancer drug screening research.

Description

technical field [0001] The invention relates to a method for culturing bladder cancer organoids in vitro, and belongs to the technical field of in vitro culture of organoids. Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system, and the incidence of bladder cancer in my country has been increasing year by year in recent years. Many patients with locally advanced or metastatic bladder cancer have unsatisfactory treatment outcomes, and some patients will change regimens or even stop medication because they cannot bear the side effects of chemotherapy. Targeted therapy, as a new treatment model with definite curative effect and good patient tolerance, has emerged in the treatment of solid malignant tumors. However, the preliminary screening of bladder cancer targeted drugs still lacks efficient and accurate in vitro models for drug screening. [0003] Tumor organoids are an in vitro three-dimensional tumor cell culture technolo...

AI Technical Summary

Problems solved by technology

Although it has been reported that the sources of successfully cultured tumor organoids include breast cancer, colon cancer, lung cancer, and esophageal cancer, among which the success rate of breast cancer and colon cancer organoids is as high as 80%, but the successful culture of bladder cancer organoids has been reported. Rarely, with a low success rate (less than 30%)

In addition, although the existing organoid culture technology can maintain the high heterogeneity of tumor samples, it still lacks the fidelity of the tumor microenvironment. For example, the existing tumor organoids do not contain the tumor microenvironment Immune cells, resulting in the cultured organoids not suitable for the screening of tumor immune drugs

Images