CAR-NK cell, and preparation method and application thereof

A cell and cell culture technology, applied in the field of CAR-NK cells and their preparation, can solve the problems of instability and transduction efficiency less than 10%, and achieve high stability, best industrial application and clinical application value, and transfection efficiency stable effect
CN108300698AActive Publication Date: 2018-07-20深圳市沃英达生命科学有限公司
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
深圳市沃英达生命科学有限公司
Publication Date
2018-07-20

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Abstract

The invention relates to a CAR-NK cell, and a preparation method and an application thereof. The preparation method comprises the following steps: S1, synthesizing a CAR sequence; S2, integrating theCAR sequence into a lentiviral target vector plasmid to obtain a target plasmid; S3, mixing the target plasmid with a 293T cell, performing culturing, collecting the obtained virus liquid, and concentrating the virus liquid to obtain the virus concentrate of the target plasmid; S4, collecting autologous plasma and peripheral blood mononuclear cells (PBMC), adjusting the cell density with an activation culture medium, performing cell culturing, transferring obtained cells into a new cell culturing bag, continuing culturing, adding a proliferation culturing medium, and obtaining the NK cell after the culturing is finished; and S5, adding the virus concentrate of the target plasmid to the NK cell, and performing incubation to obtain the CAR-NK cell. The CAR-NK cell has very strong factor secretion ability and tumor inhibition ability, and the transfection method for preparing the CAR-NK cell is efficient and stable.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a CAR-NK cell and its preparation method and application. Background technique

[0002] Natural killer cells (Nature killer cells, NK cells) are an important natural immune system lymphocytes in the body. Its phenotype is generally CD3-CD16+CD56+, mainly derived from bone marrow CD34+ lymphocytes. NK cells are not limited by MHC and can recognize and kill tumor cells without antigen sensitization. At the same time, NK cells can secrete a variety of cytokines to regulate the acquired immune response, and are a bridge connecting the innate immune response and the acquired immune response, so NK cells play an important role in the immune response against tumors and viral infections, known as The first line of defense for human health.

[0003] The recognition of tumor cells by NK cells mainly depends on the receptors on the cell surface and is not restricted by MHC. After recognizin...

Claims

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