In-vitro pure culture method for natural killer cells

Abstract

The invention relates to an in-vitro pure culture method for natural killer cells. The method includes the following steps that peripheral blood is collected, single karyocyte is separated, magnetic beads combined with specific antibodies are adopted to remove the NK cells, and NK cell populations are obtained; the NK cell populations are put into a culture bottle coated with multiple antibodies,and are subjected to activation culture with a complete medium containing IL-2, IL-7, IL-15, anti-CD16, OK432 and inactivated autologous serum, and inactivated NK cell populations are obtained; the inactivated NK cell populations are put into a culture bottle free of antibody coating and continue to carry out multiplication culture, and the natural killer cells are obtained. Any heterologous serumingredients and trophoblast cells are not used in the in-vitro pure culture method, and high amplification efficiency can be wholly obtained; meanwhile, the purity of the amplified NK cells reaches 90% or above, in-vitro efficient pure culture is achieved, and the clinical application requirement is conveniently met.

Description

technical field [0001] The invention relates to the field of cells, in particular to a method for in vitro pure culture of natural killer cells. Background technique [0002] Natural killer cells (natμral killer, NK) is a kind of human immune cells, derived from bone marrow lymphocytes. In the human body, NK cells are mainly distributed in peripheral blood and spleen, and at the same time, a small amount exists in other tissues such as lymph nodes. NK cells mainly exist in peripheral blood and spleen, but the distribution ratio varies among different populations, accounting for roughly 5% to 15% of peripheral blood lymphocytes. [0003] NK cells are generally identified and screened by the protein molecules expressed on their surface. Since the cells express CD56 and lack CD3, the marker combination CD3-CD56+ is usually used to define NK cells. In addition, NK cells also have surface molecular markers such as CD-16, CD-2, and NKP46 (Hadad et al., Front Immμnol., 2015, 6:4...

AI Technical Summary

Problems solved by technology

In this method, since the initial PBMC cells are a cell mixture with NK cell content of only 5-15%, contamination of other types of cells cannot be avoided during the subsequent culture process. Generally, the content of NK cells in the final culture is only 30-70%.

In addition, there is also a method of using trophoblast cells to stimulate the growth of NK cells. However, since the trophoblast cells are generally inactivated cancer cells in this method, the safety risk of clinical use is relatively high.

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