Culture and cryopreservation method for prostatic cancer organoid with tumor immune microenvironment

Abstract

The invention provides a culture and cryopreservation method for a prostatic cancer organoid with a tumor immune microenvironment. The culture and cryopreservation method comprises the following steps: separating mononuclear cells and autoserum from peripheral blood, and stimulating the mononuclear cells by using a CD28 antibody and IL-2; separating tumor cells from a prostatic cancer tissue sample; mixing the tumor cells with a culture medium and a temperature-sensitive hydrogel, and then carrying out culturing and subculturing to obtain a tumor organoid precursor; stimulating the tumor organoid precursor by using IFN-gamma, and then conducting dissociating to form stimulated tumor cells; mixing the stimulated tumor cells with the stimulated mononuclear cells, conducting stimulating witha PD-1 antibody, mixing the stimulated mixture with the temperature-sensitive hydrogel, and carrying out culturing; and cryopreserving the tumor organoid with a cryopreservation solution containing FBS and the temperature-sensitive hydrogel. With the method in the invention, the success rate of culturing of organoids with a tumor microenvironment can be greatly improved at low cost.

Description

technical field [0001] The present application relates to the technical field of biomedicine, and specifically relates to a method for culturing and freezing prostate cancer organoids with a tumor immune microenvironment. Background technique [0002] Tumor organoids (Organoids) are cell cultures derived from patient tumor tissue that can be grown in three dimensions (3D) in vitro. Tumor organoids preserve tumor heterogeneity, tissue characteristics, and gene mutation information, which can simulate the occurrence and development of cancer in vitro, construct disease models, and conduct fast and accurate cancer drug screening, effectively supplementing existing animal and two-dimensional The (2D) cell culture model provides a new technical platform for the rapid and effective development of tumor drugs. [0003] Prostate cancer is currently the second most deadly tumor among male malignant tumors. The current conventional treatment for advanced prostate cancer is anti-andro...

AI Technical Summary

Problems solved by technology

However, the number of prostate tumor cell lines successfully constructed so far is less than 10, which cannot represent a huge spectrum of clinical diseases

Coupled with the establishment of immortalized tumor cell lines, tumor heterogeneity and the lack of tumor immune microenvironment, the preclinical data and the real situation of patients are seriously deviated, and a large number of clinical trials of new drugs fail

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