Method for efficiently amplifying natural killer cells from peripheral blood in high purity manner

A natural killer cell and peripheral blood technology, which is applied in the field of high-efficiency and high-purity expansion of natural killer cells, can solve the problems of unresolved safety hazards, low purity, and cumbersome methods and processes, and achieves optimization of activation and amplification conditions and simplified operations. Process, enhance the effect of cytotoxicity
CN107326008AActive Publication Date: 2017-11-07SHANGHAI LIFE SCI & TECH CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
SHANGHAI LIFE SCI & TECH CO LTD
Publication Date
2017-11-07

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Abstract

The invention relates to a method for efficiently amplifying natural killer cells from peripheral blood in a high purity manner. The method comprises the following steps: collecting 100ml-200ml of peripheral blood, separating mononuclear cell, putting the mononuclear cell into a culture bottle coated with CD16 monoclonal antibody and CD226 monoclonal antibody for culturing, and activating the cell by virtue of IFN-gamma; after the activation is carried out for 24 hours, adding IL-2, IL-15, IL-21, OK432, CD2 monoclonal antibody and inactivated autoserum into a culture medium; transferring the cell into a culture bottle which is not coated with CD16 monoclonal antibody and CD226 monoclonal antibody for culturing, and supplementing liquid to the cell once in each 2-3 days; and finally transferring the cell into a culture bag for culturing, and collecting NK cell in the 21th day. According to the method, no any heterologous serum component or trophoblast cell is used, so that the safety and reliability of the prepared NK cell are guaranteed; and the method has good clinical application values.
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Description

technical field

[0001] The invention belongs to the field of cell differentiation and culture, and in particular relates to a method for amplifying natural killer cells with high efficiency and high purity from peripheral blood. Background technique

[0002] Natural killer cells (natural killer, NK) are important innate immune cells of the body, differentiated from bone marrow hematopoietic stem cells, and their development and maturation depend on the bone marrow or thymus microenvironment; mature NK cells are mainly distributed in peripheral blood and spleen, and in lymph nodes and other organizations also exist in small quantities. NK cells are larger and contain cytoplasmic granules, so they are called large granular lymphocytes. Because they do not express specific antigen recognition receptors, NK cells are the third type of lymphocytes different from T and B lymphocytes. NK cells express CD56 and CD16, but lack CD3, usually with marker combination CD3 - CD56 + to i...

Claims

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