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A kind of immature dendritic cell culture fluid and its immature dendritic cell preparation method

A dendritic cell, immature technology, used in cell culture active agents, biochemical equipment and methods, animal cells, etc., can solve the problems of slow maturation time of DC cell culture, low cell number and purity, etc.

Active Publication Date: 2021-02-12
蓝莲(杭州)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many existing DC cell culture methods, but generally there are problems such as the slow maturation time of DC cell culture, which takes about 4 days to mature, and the number and purity of cells are low.

Method used

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  • A kind of immature dendritic cell culture fluid and its immature dendritic cell preparation method

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Experimental program
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Effect test

Embodiment 1

[0031] DC culture steps

[0032] 1. Confirm that the suction tube of the plasma bag is sealed. Wipe the pipette and scissors with two different sterile room wipes soaked in 70% ethanol.

[0033] 2. Hold the suction tube with tweezers, cut the front end of the tube and control the flow rate with tweezers to transfer the plasma into a 50ml test tube. Record the plasma volume transferred.

[0034] 3. Centrifuge the tube (400g, 4°C) for 30 minutes to remove platelets.

[0035] 4. Pour the supernatant liquid into a new 50ml test tube.

[0036] 5. Put the test tube into a constant temperature water tank (56°C) and incubate for 30 minutes.

[0037] 6. Centrifuge tubes (400g, 4°C) for 60 minutes to remove precipitated fibrin.

[0038] 7. Pour the upper liquid (autologous serum) into a new 50ml test tube. The tubes were then labeled with the patient's name, date of preparation, and refrigerated (4°C).

[0039] 8. Confirm that the suction tube of the PBMC bag is sealed. Hang the...

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Abstract

The invention relates to an immature dendritic cell culture medium and a preparation method thereof, belonging to the technical field of cell culture. The cell preparation includes the following steps: 1) collecting leukocytes after processing the plasma; 2) preparing DC culture flasks for culturing PBMCs; 3) adding DC culture fluid to the flasks after culturing, and culturing for 3 days; 4) harvesting DC cells. The above-mentioned immature dendritic cell culture medium and the method for preparing the immature dendritic cell, wherein IL-13 interleukin is added to the cell culture medium, can induce monocyte differentiation and enhance the expression of MHC class II molecules; Adding anti-CD83 antibody can effectively improve the purity of immature DC cells; when harvesting DC cells, EDTA is used to relieve the cell adhesion state and improve cell survival rate; using this method, DC cell culture matures quickly, and it takes only 3 days to mature , and the cell number and purity are high.

Description

technical field [0001] The invention belongs to the technical field of cell culture, in particular to a culture solution of immature dendritic cells and a preparation method of the immature dendritic cells. Background technique [0002] Dendritic cells (Dendritic Cells, hereinafter referred to as DC) are a kind of macrophages and also the most powerful antigen presenting cells (Antigen Presenting Cells, APCs). In the human body, DC cells mainly activate helper T cells and B cells by phagocytizing antigens and expressing a part of them on MHC II molecules on the surface of DC cells. DCs have been shown to be the only ones capable of significantly stimulating naive T cells ( T cells) proliferated APC. There are many existing DC cell culture methods, but generally there are problems such as slow maturation time of DC cell culture, which takes about 4 days to mature, and low cell number and purity. Contents of the invention [0003] In view of the above-mentioned problems ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2501/22C12N2501/2304C12N2501/2313C12N2501/599
Inventor 杨罕闻
Owner 蓝莲(杭州)生物科技有限公司
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