Preparation method used for high efficiency amplification of natural killer cells

Abstract

The invention relates to a preparation method used for high efficiency amplification of natural killer (NK) cells. The preparation method comprises following steps: peripheral blood is collected, andsingle karyocytes are separated; the karyocytes are introduced into a culture bottle treated via coating with CD16 monoclonal antibody and HER2 monoclonal antibody for culturing; IL-2, IL-15, IL-21, and inactivated autologous serum are introduced into a culture medium at the same time; the cells are transformed into a culture bottle free of coating treatment, liquid supply is carried out every 2 to 3 days; and at last the obtained NK cells are collected. According to the preparation method, no heterogenous serum or trophoblastic cell is adopted, operation process is simple, high NK cell yieldis achieved, and the clinical application value is high.

Description

technical field [0001] The invention belongs to the field of cell differentiation and culture, in particular to a preparation method for efficiently expanding natural killer cells. Background technique [0002] Natural killer cells (natural killer, NK) are derived from bone marrow lymphoid stem cells. Their differentiation and development depend on the microenvironment of bone marrow or thymus. They are mainly distributed in peripheral blood and spleen, and also exist in a small amount in lymph nodes and other tissues. NK cells do not express specific antigen recognition receptors, and are the third type of lymphocytes different from T and B lymphocytes. In the process of killing target cells, NK cells can non-specifically recognize target cells and are not restricted by the major histocompatibility complex (MHC), and can rapidly kill and dissolve various tumor cells. NK cells express CD56 but lack CD3 and are usually defined by the marker combination CD3-CD56+. NK cells m...

AI Technical Summary

Problems solved by technology

However, there are many problems to be solved in the conventional method for preparing NK cells: 1. The purity of the obtained NK cells is low, and the yield is not high. The yield of NK cells from different individuals varies greatly, and the blood cells of some individuals are difficult to expand NK cells

2. If trophoblast cells are used in the process of culturing NK cells, the purity and yield of NK cells can be significantly improved, but the use of trophoblasts will greatly reduce the safety of clinical applications

However, the method of immunomagnetic beads is cumbersome and costly, and may introduce foreign substances (magnetic beads, components in the eluent, antibodies, etc.)

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