A method for isolation and induction of tumor-infiltrating T-lymphocytes suitable for malignant solid tumors

Abstract

The present invention provides a method for separating and inducing tumor-infiltrating T lymphocytes suitable for malignant solid tumors, specifically, including 1) processing the tumor tissue into a volume of 0.5-3mm 3 2) Matrigel is spread in the cell culture plate, and the tissue piece of the tumor tissue processed in step 1) is transferred to Matrigel for coating; 3) The coated tissue in step 2) Transfer the tissue block to another cell culture plate and incubate for 10-60 minutes; 4) add pre-amplification medium to the cell culture plate in step 3), and carry out pre-amplification culture for 10-30 days, every 2- For 3 days, half of the pre-amplification medium was used to change the medium. The invention provides an effective method for separating and inducing tumor infiltrating T lymphocytes for the specific biological treatment of malignant solid tumors. The tumor infiltrating T lymphocytes separated by the method of the invention have high purity and stronger tumor cell killing activity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for separating and inducing T lymphocytes infiltrated by malignant solid tumors. Background technique [0002] At present, the main treatment methods for tumor treatment include traditional radiation therapy, surgical tumor resection, drug therapy and emerging tumor biological therapy. Since surgery, radiotherapy and chemotherapy will bring huge toxic and side effects to patients, biological therapy has become the fourth important tumor treatment mode due to its significant efficacy advantages. Tumor biological therapy mainly includes: adoptive cell therapy, cytokine therapy, tumor vaccine, targeted molecular therapy, etc. At present, there are many studies on adoptive cell therapy, including tumor-infiltrating lymphocytes (TILs), which are mainly CD8 accumulated in tumor lesions. + T lymphocytes are at the forefront of immune response and regulation in tumor immune mechan...

AI Technical Summary

Problems solved by technology

These classic methods for isolating tumor-killing activity of TIL cells need to isolate and induce primary tumor cells as target cells from patient tumor tissues, and co-cultivate TIL cells and target cells in vitro (for example, 24 hours), which takes a long time. ; and isolated and cultured primary tumor cells from tumor tissue and local tumor infiltrating CD8 + T lymphocytes are also quite difficult. Therefore, it is urgent to establish an efficient and easy method to isolate and identify tumor infiltrating T lymphocytes in patients, so as to be applied to tumor immunotherapy

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