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Method for establishing NK and/or T cell line

A cell line and nuclear cell technology, applied in the field of establishing NK and/or T cell lines, can solve the problems of limiting the pathogenic mechanism of diseases, the difficulty of establishing NK cell lines and T cell lines, and the difficulty in obtaining children's patients. efficiency effect

Inactive Publication Date: 2018-07-06
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Purification of a certain population of lymphocytes requires 10 mL of whole blood, which is not readily available in pediatric patients
[0004] NK cell lines and T cell lines are not easy to establish, which limits the research on the pathogenic mechanism of related diseases, drug screening work, and the possible therapeutic application of genetically modified cell therapy to such diseases

Method used

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  • Method for establishing NK and/or T cell line
  • Method for establishing NK and/or T cell line
  • Method for establishing NK and/or T cell line

Examples

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Comparison scheme
Effect test

Embodiment 1

[0041]This embodiment is used to illustrate the method for establishing NK and / or T cell lines from peripheral blood mononuclear lymphocytes of patients with EBV infection-associated lymphoproliferation, lymphocytoma, and leukemia in the present invention, which specifically includes the following steps:

[0042] 1. Anticoagulated peripheral blood was collected from patients with lymphoproliferative disease, centrifuged in Ficoll (Ficoll-plaque TM PLUS, GE HealthCare) at 2500rpm for 30min, and peripheral blood mononuclear cells (PBMCs) were separated from the buffy coat.

[0043] 2. Resuspend the peripheral blood mononuclear cells isolated above with PBS, take 18 μL of the cell suspension and add 2 μL of trypan blue to stain for 2 minutes, count the cells with a cell counter or count the cells with a red blood cell counting board.

[0044] 3. Centrifuge at 1200rpm for 10 minutes, resuspend the cells with medium to make the concentration 1~2×10 6 / mL, add 100-200U / mL cytokines,...

Embodiment 2

[0058] The difference between this example and Example 1 is that the peripheral blood is collected from a patient with EBV infection-related lymphoma.

Embodiment 3

[0060] The difference between this example and Example 1 is that the peripheral blood is collected from a leukemia patient associated with EBV infection.

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Abstract

The invention discloses a method for establishing a cell line, and discloses a method for establishing a NK and / or T cell line. The method is specifically as follows: collecting peripheral blood mononuclear cells of patients with chronic active EBV infection or other EBV-infection-related lymphocyte proliferation, lymphoma or leukemia, and culturing the peripheral blood mononuclear cells in a medium containing human serum in the presence of cytokines. According to the method, NK and or T cells which can proliferate continuously can be established from a small amount of the peripheral blood mononuclear cells, and the obtained cell line can be continuously cultured for more than 3 months. The method requires only the small amount of the peripheral blood mononuclear cells, does not require use of expensive equipment and high concentrations of the cytokines, and can significantly improve the efficiency of establishing of cell lines.

Description

technical field [0001] The present invention relates to a method for establishing cell lines, in particular to a method for establishing NK and / or T cell lines. Background technique [0002] EBV is a tumor virus that can latently infect B cells, NK and T cells (CD4+, CD8+, γδT), etc., causing lymphoproliferation. Among them, NK and T cells play an important role in the immune system. NK cells are a kind of natural killer cells, which can kill pathogen-infected cells and cancer cells in a non-specific way, and can kill pathogens with enzymes such as perforin and granzyme, or kill pathogens through Fas-FasL interaction. Different T cells perform different functions in the immune system, including helper T cells and killer T cells. The pathogenesis of lymphoproliferative disease caused by EBV infection of immune cells is still unclear. It may be that the infected cells lose their ability to kill and cause disease, or lymphocyte proliferation may cause lymphoma and blood cance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12Q1/02A61K35/17A61P35/00A61P35/02A61P31/22
CPCA61K35/17C12N5/0636C12N5/0646C12N2501/2302C12N2501/2304C12N2501/2315C12N2501/2323G01N33/5017G01N33/505
Inventor 段子渊许崇凤谢正德张庆勋
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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