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

A technology of cell preparation and application of lymphocytes, applied in the field of CIK cell preparation and expansion of CIK, can solve the problems of unsatisfactory cell multiplication and cytotoxic activity of CIK cells, and achieve strong mitogenic effect, enhanced activation effect, and increased quantity. Effect

Active Publication Date: 2013-05-22
SHANGHAI CLAISON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 embodiment is to make those skilled in the art fully understand the technical scheme of the present invention and the corresponding technical effects by comparing with the prior art implementation. As for the various reagents used in this embodiment, they can be Reagents purchased from different manufacturers will not cause differences in the experimental results of this example after the reagents produced by different manufacturers are processed according to the routine in the field. The serum-free medium used is a commercial serum-free medium supplemented with 2% autologous serum.

[0029] (1), the method for amplifying CIK adopted in this embodiment

[0030] A method of expanding CIK comprising:

[0031] ①. Use a blood cell separator or obtain peripheral blood mononuclear cells (PBMC) from peripheral whole blood, suspend PBMC with serum-free medium, and adjust the concentration to 1×10 6 / ml, the culture bag was 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 invention belongs to the field of in vitro culture of immune cells, and relates to a method for amplifying CIK, wherein the CIK cells are cytokine-induced killer cells through expansion and culture of peripheral blood mononuclear cells (PBMC). A CIK cell preparation is provided. Background technique [0002] Biotherapy is to achieve clinical therapeutic effects by mobilizing the host's natural defense mechanism or giving naturally occurring highly targeted substances. An important method in the field of biotherapy is immune cell therapy, which is to pass the isolated autoimmune 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 reinfused into the body to play a therapeutic role. The types of such immune cells include lymphokine-activated killer cells (LAK), tumor-infiltrating lymphocytes (TIL), cytotoxic lymphocytes (CTL), CD3 mAb-activated...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/078C12N5/0786A61K35/14A61P35/00A61K35/15
Inventor 张赞苏国新
Owner SHANGHAI CLAISON BIOTECH
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