Method for simultaneous and efficient amplification of CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells

Abstract

The invention discloses a method for simultaneous and efficient amplification of CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells. The method comprises the steps as follows: the concentration of separated PBMC (peripheral blood mononuclear cells) is adjusted by a serum-free medium containing autologous plasma, an Anti-CD16 antibody, IL-2 and IL-15 are added, and then the mixture is transferred into a T175 culture flask for culture; an Anti-CD3 antibody and an Anti-CD137 antibody are added; a serum-free medium containing the autologous plasma, IL-2 and IL-15 is supplemented every two days according to the cell growth condition; the cell concentration is controlled to be about 1.5*10<6> / ml; and after culture is performed for 14-21 days, large quantities of high-purity CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells can be obtained simultaneously, and the total cell quantity can reach an effective value of the cell quantity required for adoptive cellular immunotherapy clinically for tumor. The method for simultaneous and efficient amplification of the CD<3+>CD<56+>CIK cells and the CD<3->CD<56+>NK cells is simple, convenient, effective and high in cell killing activity.

Description

technical field [0001] The invention belongs to the field of in vitro culture of immune cells, and relates to a method for simultaneously and efficiently amplifying CD from peripheral blood mononuclear cells (PBMC) 3+ cd 56+ CIK cells and CD 3- cd 56+ NK cell approach. Background technique [0002] Malignant tumor is a disease that seriously endangers human health, and its mortality rate is increasing year by year. Traditional tumor treatment methods mainly include surgery, chemotherapy and radiotherapy. However, traditional treatment methods lack specificity in killing tumor cells and damage normal tissue cells, accompanied by obvious toxic and side effects. In recent years, with the development of immunology and molecular biology techniques, the fourth mode of tumor treatment, that is, biological therapy, has emerged. Biological therapy can kill tumor cells more specifically without damage to normal tissue cells, and can activate the patient's own immune system to ge...

AI Technical Summary

Problems solved by technology

[0007] Due to the strong anti-tumor activity of CIK and NK cells, in recent years, with the advancement of immune cell preparation technology and the improvement of people's awareness and acceptance of immune cell therapy, CIK and NK cells have been widely used in clinical tumor treatment. However, the CIK and NK immune cells of the patient are expanded separately in vitro, the large amount of blood collected, and the high cost have become bottlenecks in the development of various cellular immunotherapies.

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