DC (dendritic cell) inducer and application thereof

An inducer, GM-CSF technology, applied in cell culture active agents, microorganisms, blood/immune system cells, etc., can solve the problems of low induction maturation rate, low cell proliferation rate, poor antigen presentation performance, etc. Cell viability, increase the proliferation rate and maturation rate, and improve the effect of antigen presentation

Inactive Publication Date: 2016-06-15
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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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 to provide a dendritic cell that can improve the maturation rate of dendritic cells in vitro, aiming at the defects of low in vitro induction maturation rate, low cell proliferation rate, and poor antigen presentation performance of the dendritic cells cultured in the prior art. DC Inducers with Efficiency and Antigen Presentation Performance and Their Application in Inducing DC Maturation

Method used

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  • DC (dendritic cell) inducer and application thereof
  • DC (dendritic cell) inducer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The application of the DC inducer provided by the present invention in the method for inducing DC maturation comprises the following steps:

[0033] S1a, obtaining dendritic cell precursor mononuclear cells;

[0034] Draw 3-5ml of peripheral venous blood, dilute it with normal saline 1:1, then slowly add it to 3-5ml of Ficoll-Hypaque lymphocyte separation medium with a specific gravity of 1.077±0.001g / ml along the tube wall, and centrifuge at 1500r / min for 15 minutes , isolate and obtain peripheral blood mononuclear cells, and suspend the cells in RPMI1640 medium (purchased from Gibco) containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 U / mL streptomycin at a concentration of 4×10 6 / ml, transferred to a six-well plate, 2ml / well, put the six-well plate at 37°C, 5% CO 2 Incubate in an incubator for 2 hours, remove the medium and suspended cells, put fresh RPMI1640 medium in each well, blow gently to blow up the cells on the wall, and collect the cell suspens...

Embodiment 2

[0042] The difference from Embodiment 1 is that in this embodiment,

[0043] Among the first inducers used,

[0044] The concentration of rhGM-CSF is 800U / ml,

[0045] The concentration of GM-CSF is 5ng / mL,

[0046] The concentration of IL-4 is 5ng / mL,

[0047] The concentration of rhIL-4 is 800U / ml,

[0048] and IL-13 at a concentration of 5 ng / mL;

[0049] In the second inducer,

[0050] The concentration of rhTNF-α is 800U / ml,

[0051] The concentration of gallic acid was 5 μg / ml,

[0052] Or the concentration of walnut green husk extract is 10μg / ml.

Embodiment 3

[0054] The difference from Embodiment 1 is that in this embodiment,

[0055] Among the first inducers used,

[0056] The concentration of rhGM-CSF is 1200U / ml,

[0057] The concentration of GM-CSF is 15ng / mL,

[0058] The concentration of IL-4 is 15ng / mL,

[0059] The concentration of rhIL-4 is 1200U / ml,

[0060] and IL-13 at a concentration of 15 ng / mL;

[0061] In the second inducer,

[0062] The concentration of rhTNF-α is 1200U / ml,

[0063] The concentration of gallic acid is 15μg / ml,

[0064] Or the concentration of walnut green bark extract is 30μg / ml.

[0065] In order to further verify that the DC inducer provided by the present invention and the obtained dendritic cells have significant beneficial effects, the following experiments are set up for verification.

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Abstract

The present invention relates to a DC inducer, comprising: rhGM-CSF, GM-CSF, IL-4, rhIL-4 and IL-13. The present invention further relates to the application of the above-mentioned DC inducer in the process of inducing DC maturation. Compared with the prior art, the DC inducer provided by the present invention can increase the cell activity of dendritic cells, increase the proliferation rate and maturation rate of dendritic cells, and further improve the antigen presentation of dendritic cells.

Description

technical field [0001] The invention relates to the field of tissue engineering, in particular to a DC inducing agent and its application. Background technique [0002] Cellular immunotherapy is an emerging and brand-new anti-tumor treatment method with significant curative effect. It makes up for the disadvantages of traditional surgery, radiotherapy and chemotherapy. It has been recognized as the most active and effective tumor comprehensive treatment model in the 21st century. It is a promising treatment method, and it is also the only treatment method in the world that has the hope of completely eradicating tumor cells. [0003] Cellular immunotherapy is to collect the body's own immune cells, cultivate them in vitro, increase their number by thousands of times, and enhance their targeted killing function, and then reinfuse them back into the human body to kill pathogens, cancer cells, and mutations in blood and tissues. cells, break immune tolerance, activate and enhan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2501/22C12N2501/2304C12N2501/2313
Inventor 曾宪卓鲁菲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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