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CIK (cytokine-induced killer) cell amplification method

A cell and nuclear cell technology, applied in the direction of animal cells, vertebrate cells, cell culture active agents, etc., can solve the problems of low cell killing activity and poor amplification efficiency, so as to improve the killing activity, increase the number, and improve the amplification The effect of efficiency and maturity

Inactive Publication Date: 2016-06-22
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a method that can improve the expansion efficiency of CIK cells, especially CD3 + and CD56 + The number of CIK cells, the CIK cell expansion method that improves killing activity

Method used

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Examples

Experimental program
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Embodiment 1

[0031] The CIK cell expansion method provided by the invention comprises the following steps:

[0032] S1a, mononuclear cells are isolated from peripheral blood;

[0033] S11a. Collect 20ml of peripheral blood from healthy volunteers under sterile conditions, anticoagulate with heparin, centrifuge at 20°C for 1000g×10min in a GMP clean laboratory, draw the upper layer of plasma and place it in another sterile 50ml centrifuge tube. After centrifugation, 56 Place in a water bath at ℃ for 30 minutes for later use, add D-PBS buffer and dilute the blood cell layer at a ratio of 1:1, and mix by pipetting.

[0034] S12a, using Ficoll density gradient centrifugation to separate human peripheral blood mononuclear cells;

[0035] Use two 50ml centrifuge tubes to fill 25ml of lymphocyte separation solution, and use a sterile pipette to slowly add the diluted blood to the upper layer of the lymphocyte separation solution along the wall of the centrifuge tube to form a clear interface and...

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Abstract

The invention relates to a CIK cell expansion method, comprising the following steps: obtaining mononuclear cells, using a CIK cell inducer to induce and culture the mononuclear cells until the CIK cells are harvested; placing the CIK cells in a serum-free medium In vitro expansion culture; wherein, the serum-free medium contains PPP, IL-2, IL-12 and IL-27. It can not only improve the expansion efficiency and maturation rate of CIK cells, but also increase the CD3 + and CD56 + Increase the number of CIK cells and increase the killing activity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a CIK cell expansion method. Background technique [0002] Cellular immunotherapy is an emerging and brand-new anti-tumor treatment method with significant curative effect. It makes up for the disadvantages of traditional surgery, radiotherapy and chemotherapy. It has been recognized as the most active and effective tumor comprehensive treatment model in the 21st century. It is a promising treatment method, and it is also the only treatment method in the world that has the hope of completely eradicating tumor cells. Immune cells refer to cells involved in or related to immune responses, including lymphocytes, dendritic cells, monocytes / macrophages, granulocytes, mast cells, etc. [0003] CIK (cytokine-induced killer, Chinese name: [autologous cell immunotherapy] multiple cytokine-induced killer cells) is a group of heterogeneous cell populations obtained from mononuclear cells cultu...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N2500/90C12N2501/2301C12N2501/2302C12N2501/2312C12N2501/2327C12N2501/24C12N2501/515C12N2506/11
Inventor 曾宪卓曾明哲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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