A culture method for inducing NK cells in vitro after cryopreserved human pbmc recovery

Abstract

The invention relates to a culture method for inducing NK cells in vitro after cryopreserved human PBMCs are resuscitated. Including three steps of cell recovery, NK cell induction, and NK cell expansion; wherein, in the induction stage of NK cells, use CD 16 and thymosin diluted with PBS solution to coat the culture flask; take serum-free culture medium and add IL‑ 2. IL-12 and IL-15 were prepared as induction medium; serum-free culture solution was added to IL-2 and IL-7 to prepare activation medium; serum-free culture solution was added to IL-2 to prepare expansion medium, On day 3, the half-change medium was used for rehydration for activation culture. Utilizing the culture method of the present invention to induce NK cells in vitro after recovery of cryopreserved human PBMCs is simple and efficient, which not only reduces the cost of cell culture, but also improves the safety of cell culture, and can effectively overcome the difficulty of inducing NK cells from cryopreserved PBMCs and increase the number To achieve the purpose of cell therapy and enhance its anti-tumor, anti-virus and anti-infection capabilities.

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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