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Immature dendritic cell culture solution and preparation method of immature dendritic cells

A technology of dendritic cells and culture medium, applied in the field of cell culture, can solve the problems of low cell number and purity, slow maturation time of DC cell culture, etc.

Active Publication Date: 2019-08-09
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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  • Immature dendritic cell culture solution and preparation method of immature dendritic cells

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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 provides an immature dendritic cell culture solution and a preparation method of immature dendritic cells, and belongs to the technical field of cell culture technologies. The cell preparation method comprises the following steps that 1, after blood plasma treatment, leukocytes are collected; 2, a DC culture bottle is prepared, and culture of PBMCs is conducted; 3, after culture, theDC culture solution is added into the bottle, and culture is conducted for 3 days; 4, DCs are harvested. According to the immature dendritic cell culture solution and the preparation method of the immature dendritic cells, IL-13 interleukin is added into the cell culture solution, mononuclear cell differentiation can be induced, and expression of MHC-II molecules is enhanced. By adding an anti-CD83 antibody, the purity of the immature DCs can be effectively improved; when the DCs are harvested, EDTA is used to relieve the cell attachment state, the cell surviving rate is improved; by using the method, the DC culture maturation time is short, it only takes 3 days to mature the DCs, and the number and the purity of the cells 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 Applications(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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