Preparation method used for high efficiency amplification of natural killer cells

A natural killer cell, high-efficiency technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of low yield, high cost, low purity of NK cells, etc., to solve safety problems, operation Simple and stable results

Inactive Publication Date: 2018-06-01
上海莱馥生命科学技术有限公司
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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...

Method used

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  • Preparation method used for high efficiency amplification of natural killer cells
  • Preparation method used for high efficiency amplification of natural killer cells

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Embodiment 1

[0023] 1. Reagents and consumables required for cell preparation

[0024] HER2 monoclonal antibody (Herceptin, Herceptin): 440 mg per bottle, purchased from Roche Pharmaceutical Company.

[0025] CD16 monoclonal antibody: purchased from Beijing Tongli Haiyuan Biological Company.

[0026] Recombinant human interleukin 21 (IL-21): purchased from Beijing Tongli Haiyuan Biological Company.

[0027] Recombinant human interleukin 15 (IL-15): purchased from Beijing Tongli Haiyuan Biological Company.

[0028] Recombinant human interleukin 2 (IL-2): purchased from Beijing Tongli Haiyuan Biological Company.

[0029] X-VIVO TM 15 Serum-free cell culture medium: purchased from Lonza Company of the United States.

[0030] Lymphocyte separation medium: purchased from GE Healthcare, USA.

[0031] Cell culture flasks: purchased from Corning Company, USA.

[0032] 2. Preparation of NK cells

[0033]1. Coat culture flasks with anti-CD16 and HER2 monoclonal antibodies

[0034] Dilute ant...

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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...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/11C12N2501/2302C12N2501/2315C12N2501/2321C12N2501/599
Inventor 李新峰金乐汪佳莹李文荣李超
Owner 上海莱馥生命科学技术有限公司
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