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Method for amplifying cytokine induced kill cells (CIK) and CIK cell preparation

A cell preparation, technology using lymphocytes, applied in the field of in vitro culture of immune cells, can solve the problems of unsatisfactory proliferation and cytotoxic activity of CIK cells, achieve strong mitogenic effect, increase the expansion level, and increase the number of effects

Active Publication Date: 2012-02-15
SHANGHAI CLAISON BIOTECH
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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 that the CIK cell proliferation cell multiple and cytotoxic activity obtained by the conventional CIK amplification method are not ideal enough, and a method for amplifying CIK is provided for this problem

Method used

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  • Method for amplifying cytokine induced kill cells (CIK) and CIK cell preparation
  • Method for amplifying cytokine induced kill cells (CIK) and CIK cell preparation
  • Method for amplifying cytokine induced kill cells (CIK) and CIK cell preparation

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Embodiment

[0028] The purpose of this example is to enable those skilled in the art to fully understand the technical scheme of the present invention and the corresponding technical effects through comparison with the prior art implementation. As for the various reagents used in this example, all reagents are available on the market. Commercially available, reagents produced by different manufacturers are processed in accordance with routine procedures in the field, which will not cause different experimental results in this example. The serum-free medium used is a commercial serum-free medium supplemented with 2% autologous serum.

[0029] (1) The method for amplifying CIK used in this example

[0030] A method of amplifying CIK includes:

[0031] ①. Use a blood cell separator or obtain peripheral blood mononuclear cells (PBMC) from peripheral whole blood, suspend the PBMC in a serum-free medium, and adjust the concentration to 1×10 6 / ml, the culture bag is pre-coated with 10ug / ml CD3mAb and ...

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Abstract

The invention relates to a method for amplifying cytokine induced kill (CIK) cells and a CIK cell preparation, which belong to the field of in-vitro culture of immune cells. The method concretely adopts the following procedures that: a, lymphocyte cell separation liquid is used for separating out peripheral blood mononuclear cells (PBMC), a culture bag is covered by CD3mAb and CD137mAb in advance, the concentration of the PBMC obtained through separation is regulated to 1*10<6> / ml by a serum-free culture medium, in addition, IFN-gamma is added to obtain the final concentration being 1000 mu / ml, and the materials are transferred to the culture bag to be cultured; b, CD3mAb, CD28mAb and CD137mAb are added after the culture for 24h, in addition, the prepared serum-free culture medium is added, IL-1alpha, IL-2, IL-12 and IL-15 are added into the prepared serum-free culture medium, and obtained CIK cells are collected through centrifugation after the continuous culture for 7 to 21 days; and c, in the culture process of the step b, the cells in the culture bag are counted every three days, in addition, the culture medium is supplemented according to the concentration of the cells, and the CD3mAb, the CD28mAb and the CD137mAb are added to the corresponding concentration every six days, so the CIK cell generative cell times and the cytotoxin activeness are improved.

Description

Technical field [0001] The present invention belongs to the field of in vitro culture of immune cells, and relates to a method for amplifying CIK. The CIK cells are killer cells induced by cytokine expansion and culture via peripheral blood mononuclear cells (PBMC). The present invention is still further A CIK cell preparation is provided. Background technique [0002] Biological therapy is to achieve clinical therapeutic effects by mobilizing the host's natural defense mechanism or giving naturally-produced highly targeted substances. An important method in the field of biological therapy is immune cell therapy, which means that isolated autoimmune cells are passed through cells in vitro Induced by factors, a large number of heterogeneous cell populations (CIK) with high cytotoxic activity are expanded at the beginning, and then returned to the body to play a therapeutic effect. The types of immune cells include lymphokine-activated killer cells (LAK), tumor infiltrating lympho...

Claims

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Application Information

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IPC IPC(8): C12N5/078C12N5/0786A61K35/14A61P35/00A61K35/15
Inventor 张赞苏国新
Owner SHANGHAI CLAISON BIOTECH
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