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DC cell culture reagent and culture method thereof

A cell culture and culture method technology, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problem that the human immune system has little anti-tumor effect, cannot achieve antigen clearance function, and lacks activation of aggregated T cells. Cell and other issues, to achieve good cell viability and purity, good cell viability, and the effect of promoting proliferation

Active Publication Date: 2015-11-25
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It only recognizes the protection of invading organisms derived from microorganisms, and after activating DC cells, it has little effect on the anti-tumor effect of the human immune system
Moreover, the mature DC cells produced by this culture scheme are weak in the transmission of tumor antigens, and the tumor killing effect is not good after reinfusion into the human body; they lack the function of activating and gathering T cells in exogenous allergens, and they cannot achieve the effect in disease infection. Fast and efficient antigen clearance function

Method used

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  • DC cell culture reagent and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0063] Example 1 DC-CIK cell co-culture example

[0064] (1) Mononuclear cell isolation

[0065] 1. Peripheral blood or umbilical cord blood 40mL, centrifuge at 500-800g for 10 minutes, absorb the upper plasma, inactivate at 56°C for 30min, centrifuge at 2000-3000g for 5min, supernatant plasma at 2-8°C for later use;

[0066] 2. For the lower layer of blood, add 2 times the volume of normal saline to resuspend. According to the diluted blood: Ficoll separation solution ratio of 2:1, slowly add the diluted blood to the upper layer of Ficoll separation solution to make the layer clear;

[0067] 3. Centrifuge at 500-700g for 15-30min, and absorb the middle buffy coat layer, which is mononuclear cells.

[0068] (2) DC cell isolation and culture

[0069] 1. Dilute the obtained mononuclear cells with normal saline, centrifuge at 200-400g for 5min, and wash twice;

[0070] 2. Add serum-free medium (containing 5% autologous plasma) to resuspend the cells, press 2-8×10 6 / mL inocul...

Embodiment 2D

[0073] Example 2 DC-CIK cell co-culture example

[0074] (1) Mononuclear cell isolation

[0075] 1. Peripheral blood or umbilical cord blood 40mL, centrifuge at 500-800g for 10 minutes, absorb the upper plasma, inactivate at 56°C for 30min, centrifuge at 2000-3000g for 5min, supernatant plasma at 2-8°C for later use;

[0076] 2. For the lower layer of blood, add 2 times the volume of normal saline to resuspend. According to the ratio of diluted blood: Ficoll separation solution 2:1, slowly add the diluted blood to the upper layer of Ficoll separation solution to make the layer clear;

[0077] 3. Centrifuge at 500-700g for 15-30min, and absorb the middle buffy coat layer, which is mononuclear cells.

[0078] (2) DC cell isolation and culture

[0079] 1. Dilute the obtained mononuclear cells with normal saline, centrifuge at 200-400g for 5min, and wash twice;

[0080] 2. Add serum-free medium (containing 15% autologous plasma) to resuspend the cells, press 2-8×10 6 / mL inocu...

Embodiment 3

[0083] Embodiment 3DC-CIK cell co-culture example

[0084] (1) Mononuclear cell isolation

[0085] 1. Peripheral blood or umbilical cord blood 40mL, centrifuge at 500-800g for 10 minutes, absorb the upper plasma, inactivate at 56°C for 30min, centrifuge at 2000-3000g for 5min, supernatant plasma at 2-8°C for later use;

[0086] 2. For the lower layer of blood, add 2 times the volume of normal saline to resuspend. According to the ratio of diluted blood: Ficoll separation solution 2:1, slowly add the diluted blood to the upper layer of Ficoll separation solution to make the layer clear;

[0087] 3. Centrifuge at 500-700g for 15-30min, and absorb the middle buffy coat layer, which is mononuclear cells.

[0088] (2) DC cell isolation and culture

[0089] 1. Dilute the obtained mononuclear cells with normal saline, centrifuge at 200-400g for 5min, and wash twice;

[0090] 2. Add serum-free medium (containing 10% autologous plasma) to resuspend the cells, press 2-8×10 6 / mL ino...

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Abstract

The invention relates to the technical field of cell culture, and in particular relates to a DC cell culture reagent and a culture method thereof. The DC cell culture reagent includes a first DC cell culture reagent and a second DC cell culture reagent, wherein the first DC cell culture reagent consists of GM-SCF and IL-4; and the second DC cell culture reagent consists of GM-SCF, IL-4, TNF-alpha and MDC. The method disclosed by the invention, by improving the type and the concentration of cell factors in a DC cell culture process, can be used for promoting the proliferation in the quantity of DC cells and keeping a relatively good cell viability, so that the antigen presentation capacity of the DC cells is enhanced and a better antitumor effect is guaranteed.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a DC cell culture reagent and a culture method thereof. Background technique [0002] Tumor is a new organism formed by the body under the action of various tumorigenic factors, and the cells of local tissues lose their normal regulation of their growth at the gene level, resulting in abnormal proliferation and differentiation. Once a new organism is formed, it does not stop growing due to the elimination of the cause. Its growth is not regulated by normal body physiology, but destroys normal tissues and organs. This is especially obvious in malignant tumors. Compared with benign tumors, malignant tumors grow faster and show invasive growth, are prone to bleeding, necrosis, ulcers, etc., and often have distant metastasis, resulting in emaciation, weakness, anemia, loss of appetite, fever, and severe organ damage. Impairment of function, etc., eventually lead to death of the...

Claims

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Application Information

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IPC IPC(8): C12N5/0784
Inventor 陈海佳王一飞葛啸虎应杰张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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