Method for preparing NK (natural killer) cell

Abstract

The invention provides a method for efficiently preparing an NK cell, which can improve the multiplication rate and the purity of the NK cell through combination of stimulation effects of cell factors and feeder cells. The method is mainly characterized by comprising the steps as follows: NCR3LG1 and m IL-15 are transfected to a K562 cell simultaneously, m IL-15 can be used for adjusting activation and multiplication of the NK cell, NCR3LG1 serving as a ligand of NKp30 which is one of main activated receptors on the surface of the NK cell can effectively stimulate activation of the NK cell, and NCR3LG1 and m IL-15 have a synergistic effect; and PBMC(peripheral blood mononuclear cells) can be multiplied over 500 times in 21 cultivation days through stimulation of factors of freely added IL-2, IL-21 and the like, and a proportion of CD3-CD56+NK cell exceeds 70%; and up to now, a research report that NCR3LG1 and m IL-15 are transfected to the feeder cells simultaneously and jointly stimulate and activate the NK cell in combination of free cell factors is absent. The invention firstly provides a method for jointly cultivating and preparing the NK cell in combination of the feeder cells and the free factors.

Description

technical field

[0001] The invention belongs to the field of cell culture, in particular to a natural killer cell (Natural Killer, NK) culture, that is, using transgenic feeder cells and cytokines to jointly stimulate peripheral blood mononuclear cells (Peripheral Blood Mononuclear Cell, PBMC) to obtain a large number of high Purity of NK cells. Background technique

[0002] NK cells are a group of larger granulated lymphocytes that express CD56 on the cell surface and lack CD3 expression, so the surface marker CD3 is usually used ‐ CD56 + to define NK cells. NK cells can account for about 5-15% of peripheral blood lymphocytes, and are distributed in the liver, viscera, bone marrow, and lymph nodes of the human body.

[0003] NK cells are an important part of the innate immune system, and can kill tumor cells or infected cells in a variety of ways: 1. After contacting with target cells, they release perforin and granzyme; 2. Through their own expressed ligands, Stimulate...

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