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Method for evaluating effectiveness of chimeric antigen receptor-T (CAR-T) cells

An effective and cellular technology, applied in the field of CAR-T cell preparation, which can solve the problems of inaccuracy, low success rate of inoculation, and complicated operation.

Pending Publication Date: 2021-03-26
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The first purpose of the present invention is to provide a method for evaluating the effectiveness of CAR-T cells, which solves the problems of inaccuracy due to lack of tumor characteristics, complicated operation, and low success rate of inoculation in the prior art; The co-culture of organoids and CAR-T cells, detection of relevant cytokines and microscope observation of the interaction between tumor organoids and CAR-T cells and the changes in cell morphology after killing can more intuitively and comprehensively confirm the effect of CAR-T cells on tumors. Whether the organoid has a killing effect, because the tumor organoid has tumor characteristics similar to the tumor tissue of the patient, and does not require the construction of an animal model, the method is more accurate, comprehensive, intuitive and simple

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  • Method for evaluating effectiveness of chimeric antigen receptor-T (CAR-T) cells

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Embodiment Construction

[0010] In a first aspect, the present invention provides a method for evaluating the effectiveness of CAR-T cells.

[0011] A method for evaluating the effectiveness of CAR-T cells, comprising the steps of:

[0012] (a) The tumor organoids were digested, cell stained a, and cell stained b CAR-T cells were resuspended and then co-cultured to obtain a co-culture system;

[0013] (b) taking the supernatant of the co-culture system in step (a) to measure cytokines;

[0014] (c) The co-culture system was stained with a granzyme B antibody conjugated to a fluorescent dye, and the killing of tumor organoids by CAR-T cells was observed microscopically.

[0015] The tumor may include tumors such as bladder cancer, kidney cancer, or upper urinary tract urothelial cancer.

[0016] The CAR-T cells contain antibodies capable of specifically binding to specific targets on the cell surface of the tumor or tumor organoid.

[0017] In some embodiments, the tumor is bladder cancer, and the s...

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Abstract

The invention provides a method for evaluating the effectiveness of chimeric antigen receptor-T (CAR-T) cells. According to the method, by co-culturing tumor organs and the CAR-T cells, related cytokines are detected and the interaction between the tumor organs and the CAR-T cells and the morphological change of killed cells are microscopically observed to evaluate the effectiveness of the CAR-T cells, and whether the CAR-T cells have a killing effect on tumor organs or not can be evaluated more visually, accurately and comprehensively; and the method has relatively high accuracy, reproducibility and simplicity and convenience in operation.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for evaluating the effectiveness of CAR-T cells and a method for preparing CAR-T cells. Background technique [0002] With the acceleration of urbanization and aging process, malignant tumors have become the second leading cause of death in the world. Although surgery, radiotherapy and chemotherapy are still routine methods for clinical treatment and alleviation of malignant tumor disease progression. However, individual differences and drug resistance are important sources of clinical treatment failure. Tumor individualized immunotherapy has become a hot spot in medical research all over the world in recent years, especially in the field of chimeric antigen receptor (CAR)-T cells, which have made major breakthroughs in the immunotherapy of various tumors. According to the surface antigen expression profile of different patients, a CAR molecular structure specifica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N5/09C12N15/867C12N5/10
CPCG01N33/5011G01N33/505C12N5/0693C12N15/86C12N5/0636C12N2503/02G01N2500/10C12N2509/00C12N2501/11C12N2501/115C12N2501/15C12N2501/727C12N2501/155C12N2501/415C12N2500/32C12N2500/38C12N2740/15043C12N2800/107C12N2510/00
Inventor 黄卫人余磊陈巍李志超
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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