Preparation method of Mesothelin chimeric antigen receptor modified T cell with targeted high-intensity tumor killing activity

Abstract

The invention relates to a preparation method of Mesothelin chimeric antigen receptor modified T cells with targeted high-intensity tumor killing activity, which comprises the following steps: transforming MSLN-CAR-puro plasmids into escherichia coli competent cells, conducting coating, screening and culturing on a solid culture medium, and extracting plasmids after propagating a grown single colony; preparing human peripheral blood mononuclear lymphocytes (PBMCs); and extracting plasmids, mixing the extracted plasmids, Super PiggyBac Transspoase plasmids and an electrotransfection buffer solution, transfecting PBMC (peripheral blood mononuclear cells), conducting culturing by using an X-VIVO culture medium, activating and amplifying CD3 positive T cells by using CD3 / CD28 magnetic beads, and screening and amplifying the transfected T cells. The mesothelin-targeted MSLN-CAR-T cell is constructed through a PiggyBac transposon system, so that the mesothelin-targeted MSLN-CAR-T cell has targeted killing performance on solid tumors and is good in safety, and a research basis is provided for related clinical transformation.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a preparation method of T cells modified by Mesothelin chimeric antigen receptors with targeted high-intensity tumor-killing activity. Background technique [0002] As one of the diseases that seriously threaten human health, cancer not only harms patients physically and mentally, but also brings huge economic burdens to families, society and the country. At present, common clinical treatment methods for malignant tumors include surgery, radiotherapy and chemotherapy, and targeted therapy, but such treatments cannot effectively control the progression and recurrence of some malignant tumors, nor can they effectively prolong the survival of such patients and improve their quality of life. Therefore, there is an urgent need to find new treatments. [0003] In recent years, immunotherapy has become an important means of tumor treatment, bringing hope to overcome ma...

AI Technical Summary

Problems solved by technology

However, the research of this technology in solid tumors is not yet mature enough to achieve the desired therapeutic effect, and still faces many challenges, including: the special pathological characteristics of solid tumor tissues, the immunosuppressive microenvironment around the tumor, the optimal combination of co-stimulatory molecules, Design of suicide genes, adverse reactions after reinfusion, etc.

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