Culture method of dendritic cells and dendritic cells

A dendritic cell and culture method technology, applied in the field of dendritic cell and dendritic cell culture, can solve the problems of low induction maturation rate, low cell proliferation rate, poor antigen presentation performance, etc. rate and maturation rate, promoting improvement, and improving the effect of antigen presentation

Inactive Publication Date: 2016-06-01
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0006] 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. Dendritic Cell Culture Method for Efficiency and Antigen Presentation Properties

Method used

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  • Culture method of dendritic cells and dendritic cells
  • Culture method of dendritic cells and dendritic cells

Examples

Experimental program
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Effect test

Embodiment 1

[0045] The culture method of dendritic cells provided by the invention comprises the following steps:

[0046] S1a, obtaining dendritic cell precursor mononuclear cells;

[0047] 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 suspension, that is, a single dendritic cell precurs...

Embodiment 2

[0057] The difference from Example 1 is that the second inducer used in this example includes: 1000 U / ml rhTNF-α and 10 μg / ml tomato water-soluble extract.

[0058] Wherein, the preparation method of tomato water-soluble extract is:

[0059] Accurately weigh 0.5g of tomato powder in a 100mL polytetrafluoroethylene microwave extraction tube, add 25mL of ethanol extraction solution, fix the microwave extraction tube in the microwave extraction device, and connect it to a temperature sensor; set the heating program according to the experimental design, Heat to the predetermined extraction temperature of 55°C within 5 minutes. After the sample is extracted for 1 hour, it is cooled for 10 minutes, and then centrifuged at 4000 rpm for 5 minutes. Under the same conditions, the extraction residue was repeated twice, and the combined supernatant was passed through a 0.2 μm PTFE filter membrane for later use.

[0060] In order to further verify that the method for culturing dendritic c...

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Abstract

The invention relates to a culture method of dendritic cells. The culture method includes the following steps that a single karyocyte of a dendritic cell precursor is obtained, culturing is conducted for 4-7 days through a first inductive agent, and immature dendritic cells are obtained; then, a second inductive agent is added to culture the immature dendritic cells till the dendritic cells are mature; the first inductive agent comprises rhGM-CSF and rhIL-4; the second inductive agent comprises rhTNF-alpha, tomato fat-soluble extract or tomato water-soluble extract. The invention further relates to the dendritic cells obtained through the method. Compared with the prior art, the method improves cell activity of the dendritic cells, the value-added ratio and maturing rate of the dendritic cells are raised, and thus the antigen presentation property of the dendritic cells is improved.

Description

technical field [0001] The invention relates to the field of tissue engineering, in particular to a method for culturing dendritic cells and dendritic cells. 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,...

Claims

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Application Information

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IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2500/76C12N2501/22C12N2501/2304C12N2501/25
Inventor 曾宪卓鲁菲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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