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Method for efficient acquisition of DC cells

A cell-based and efficient technology, applied in the field of medical immunity, can solve the problems of insufficient number of DC cells obtained, affecting the activation of CIK cells, etc.

Inactive Publication Date: 2016-11-16
SINOBIOWAY CELL THERAPY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this technique, DC cells and CIK cells need to be cultured separately, both of which are isolated from PBMCs. The current method of isolating DCs and CIK cells from PBMCs is to incubate for half an hour, resulting in DC cells The number of acquisitions is not enough, which affects the subsequent activation of CIK cells

Method used

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  • Method for efficient acquisition of DC cells
  • Method for efficient acquisition of DC cells

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Embodiment 1

[0024] A method for efficiently obtaining DC cells proposed by the present invention comprises the following steps:

[0025] A. Human peripheral blood mononuclear cells were resuspended in GT-T551 medium and incubated for 1.5 hours;

[0026] B. After removing non-adherent cells, add DC medium (DC grow) to culture DC cells.

Embodiment 2

[0028] A method for efficiently obtaining DC cells proposed by the present invention comprises the following steps:

[0029] A. Resuspend human peripheral blood mononuclear cells in GT-T551 medium and incubate for 2 hours at a temperature of 37°C in an environment of 5% CO 2 ;

[0030] B. After removing non-adherent cells, add DC medium (DC grow) to culture DC cells.

Embodiment 3

[0032] A method for efficiently obtaining DC cells proposed by the present invention comprises the following steps:

[0033] A. Resuspend human peripheral blood mononuclear cells in GT-T551 medium and incubate for 2.5 hours. The incubation temperature is 37°C and the incubation environment is 5% CO 2 ;

[0034] Wherein, the preparation method of human peripheral blood mononuclear cells is as follows:

[0035] S1. Inject human peripheral blood into a centrifuge tube and perform a centrifugation to obtain material a;

[0036] S2. Remove the upper layer of plasma, then take out the gray-yellow layer in the middle, and then add physiological saline to dilute to obtain material b;

[0037] S3, adding material b to the upper layer of the lymphocyte separation solution, followed by secondary centrifugation, and then absorbing the buffy coat layer to obtain material c;

[0038] S4. Adding physiological saline to the material c for dilution, followed by centrifugation three times to...

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Abstract

The invention discloses a method for efficient acquisition of DC cells, wherein the method comprises the following steps: A, re-suspending mononuclear cells in human peripheral blood in a GT-T551 medium and incubating the cells for 1.5-2.5h; and B, removing non-adherent cells and cultivating the DC cells with the addition of a DC medium. In comparison with the prior art that cells are incubated for 0.5h, the method disclosed by the invention, which incubates the cells for 2h, can achieve the full sedimentation of the DC cells, and pseudopods extend and the cells adhere to the bottom of a culture flask, so that complete separation from T lymphocytes is achieved.

Description

technical field [0001] The invention relates to the technical field of medical immunity, in particular to a method for efficiently obtaining DC cells. Background technique [0002] DC-CIK biological immunotherapy (or DC+CIK) refers to CIK cells co-cultured with DC cells. Cytokine-induced killer cells (cytokine-induced killers, CIK) are a kind of anti-tumor and anti-virus effector cells, which can be induced and proliferate in large quantities in vitro. Dendritic cells (DC) are effective professional antigen-presenting cells. Mature DCs can present tumor antigens through type II histocompatibility antigen (MHC-II) and other pathways, effectively resisting the immune escape of tumor cells. mechanism. CIK cells and DC cells are two important components of cellular immunotherapy, and the combination of the two can ensure an efficient immune response. In this technique, DC cells and CIK cells need to be cultured separately, both of which are isolated from PBMCs. The current me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784
CPCC12N5/0639
Inventor 徐华栋许国贞刘振云钱鹏王庆丽周龙
Owner SINOBIOWAY CELL THERAPY CO LTD
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