Culture method for in-vitro induction of NK cell after recovery of cryopreserved human PBMC

Abstract

The invention relates to a culture method for in-vitro induction of an NK cell after recovery of a cryopreserved human PBMC. The culture method comprises three steps including cell recovery, inductionof the NK cell and NK cell proliferation, wherein in the stage of indication of the NK cell, CD 16 and thymosin are diluted with a PBS solution for culture flask enveloping; IL-2, IL-12 and IL-15 areadded to a serum-free culture solution for preparation of an induction medium; IL-2 and IL-7 are added to the serum-free culture solution for preparation of an activation culture medium; and IL-2 isadded to the serum-free culture solution for preparation of a proliferation culture medium, and half-liquid replacement is carried out on the day 3 for solution replacement and activation culture. Themethod for in-vitro induction of the NK cell after recovery of the cryopreserved human PBMC is simple and efficient, reduces the cost of cell culture and also increases safety of cell culture, can effectively overcome the difficulty in inducing the NK cell with the cryopreserved PBMC and deficiency in a proliferation amount, reaches a purpose of cell treatment and enhances resistance to tumors, viruses and infection thereof.

Description

technical field [0001] The invention belongs to the technical field of immune cell therapy, and relates to a culture method for inducing NK cells in vitro after resuscitating cryopreserved human PBMC (peripheral blood mononuclear cells). [0002] technical background [0003] Natural killer cells (NK cells), a group of large granular lymphocytes, are the body's first line of defense against pathogens and tumor cells. NK cells are mainly distributed in peripheral blood, accounting for 5-10% of peripheral lymphocytes. NK activity also exists in lymph nodes and bone marrow, but the level is lower compared with peripheral blood. [0004] With the in-depth research on NK cells, NK cells are more and more widely used in the human body. At present, they have good effects in anti-tumor, anti-aging, sub-health treatment, and disease prevention. First, NK cells have cytotoxic function, which can directly kill tumor cells. Secondly, NK cells can bind to receptors on the surface of agi...

AI Technical Summary

Problems solved by technology

In this way, the cells after cryopreservation and recovery will be affected by the preparation of the cryopreservation solution and the freezing time. Although there is no obvious difference in morphology, there will be a large loss in number, and it is difficult to induce NK cells.

In addition, the current culture methods of peripheral blood NK cells mainly include factor induction method, trophoblast cell co-cultivation method, and flow sorting cell re-cultivation method; NK cell viability is poor, the induction method is unstable and controversial

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