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Method for obtaining antigen-specific T cells

A specific and cell-based technology, applied in the field of cell biology, can solve the problems of small quantity, low purity, and difficulty in obtaining antigen-specific T cells, and achieve the effect of increasing production and improving efficiency

Pending Publication Date: 2022-04-12
杭州芯递力生物科技有限公司
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  • Abstract
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Problems solved by technology

However, the adherent cells obtained by this method still contain T cells, which directly leads to the low purity and small number of mature DC cells obtained.
And at present, mature DC cells and T cells are usually only co-cultured for one round of stimulation, and unpurified T cells are used, in which the initial T cell content is low and the purity is not high
However, since it is naive T cells that can be activated by mature DC cells and receive the antigen information presented by mature DC cells, it is difficult to co-culture mature DC cells when the content of naive T cells is low and the purity is not high. Efficiently obtain antigen-specific T cells from the cell suspension obtained after unpurified T cells

Method used

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  • Method for obtaining antigen-specific T cells
  • Method for obtaining antigen-specific T cells
  • Method for obtaining antigen-specific T cells

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Embodiment Construction

[0036] The present invention will be specifically introduced below in conjunction with the accompanying drawings and specific embodiments.

[0037] The following experiments 1 to 4 obtained antigen-specific T cells according to the method of the following examples, including the following steps:

[0038] Step 1, collect peripheral blood, separate and obtain PBMC of mononuclear cells:

[0039] The peripheral blood sample was centrifuged; the supernatant was inactivated in a water bath, cooled and centrifuged, and the supernatant was taken as autologous plasma for later use; the remaining sample was diluted with 0.9% sodium chloride injection, and mononuclear cells were obtained by separating them with lymphocyte separation medium ( PBMC);

[0040] Step 2, obtaining immature DC cells:

[0041] PBMC were centrifuged to discard the supernatant, and CD14 immunomagnetic bead separation kit was used to separate mononuclear cells (immature DC cells), each 1×10 7 Add 80 μl buffer A ...

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Abstract

The invention discloses a method for obtaining antigen-specific T cells, which comprises the following steps: step 1, collecting peripheral blood, and separating to obtain mononuclear cells PBMC; step 2, obtaining immature DC cells; step 3, stimulating and inducing immature DC cells to be mature; 4, separating and purifying the initial T cells; 5, mixing the mature DC cells and the initial T cells according to the cell quantity ratio of 1: (5-120), adding a target antigen for co-stimulation culture for 1-3 rounds, and continuing to culture for 5-128 hours to obtain a cell suspension; 6, separating and purifying antigen-specific T cells; by adopting the method, the proportion of the antigen T cells in the co-culture cell suspension can be effectively increased.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for obtaining antigen-specific T cells. Background technique [0002] At present, the commonly used method for obtaining antigen-specific T cells is mainly to separate mononuclear cells (Peripheral Blood Mononuclear Cells, PBMCs) from peripheral blood, add them to an appropriate amount of medium, and incubate them in an incubator for 2 hours. Immature DC cells are considered to be immature DC cells, and suspension cells are considered to be T cells; the suspension cells are collected and washed and stored as T cells; the target antigen is co-incubated with adherent cells (immature DC cells), and by adding a variety of cells The factors induce the maturation of immature DC cells; then the obtained mature DC cells are co-cultured with the previously collected T cells to obtain antigen-specific T cells. However, the adherent cells obtained by this method still contain T cells, ...

Claims

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Application Information

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IPC IPC(8): C12N5/0786C12N5/0783
Inventor 韩宁张凡
Owner 杭州芯递力生物科技有限公司
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