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Dendritic cell culture medium and culture method

A technology of dendritic cells and culture medium, applied in cell culture active agent, culture process, tissue culture and other directions, can solve the problems of high price, hidden dangers, small number of DCs, etc., and achieves rapid expansion, easy promotion, Well-defined effects

Inactive Publication Date: 2018-06-05
重庆斯德姆生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the amount of DC in the body is extremely small, which is difficult to meet the needs of clinical treatment research.
However, with the development of technology, people have also developed media for DC expansion in vitro, but some of these media contain animal-derived substances, which have created some safety hazards. Although some do not contain animal-derived ingredients, they use The human protein or recombinant protein is expensive, which is not conducive to large-scale promotion

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0017] A culture medium for dendritic cells, including a basic medium and additional components, wherein the basic medium consists of glucose injection, physiological saline injection, amino acid injection and trace element injection in a volume ratio of 4:3:1.5: 1 Mix, the added ingredients include the following final concentration components: GM-CSF 100ng / mL, glutamine 2mmol / L, astragalus polysaccharide 0.2mg / L, notoginseng polysaccharide 0.2mg / L, chicken embryo extract 1v / v %, ginger juice 1v / v%, kiwi fruit juice 1v / v%, IL-415ng / mL, TNF-α800UI / mL, iron citrate 300mg / L and vitamin C 3mg / L.

[0018] Among them, the astragalus polysaccharide is prepared by the following method: take the waste residue extracted from total astragalus saponins, add water 3 times the weight of the waste residue, boil for 40 minutes, filter to obtain the first filtrate and the first precipitate, and add 2 times the weight to the first precipitate water, boiled for 30min, filtered to obtain the seco...

Embodiment 2

[0024] A culture medium for dendritic cells, including a basic medium and additional components, wherein the basic medium consists of glucose injection, physiological saline injection, amino acid injection and trace element injection in a volume ratio of 6:5:2: 1.5 Mixing, the added ingredients include the following final concentration components: GM-CSF 200ng / mL, glutamine 4mmol / L, astragalus polysaccharide 0.5mg / L, notoginseng polysaccharide 0.5mg / L, chicken embryo extract 3v / v %, ginger juice 3v / v%, kiwi fruit juice 3v / v%, IL-430ng / mL, TNF-α1200UI / mL, iron citrate 500mg / L and vitamin C 6mg / L.

[0025] Among them, the astragalus polysaccharide is prepared by the following method: take the waste residue extracted from total astragalus saponins, add water 5 times the weight of the waste residue, boil for 60 minutes, filter to obtain the first filtrate and the first precipitate, and add 4 times the weight to the first precipitate water, boiled for 50min, filtered to obtain the ...

Embodiment 3

[0031] A culture medium for dendritic cells, including a basic medium and additional components, wherein the basic medium consists of glucose injection, physiological saline injection, amino acid injection and trace element injection in a volume ratio of 4.5:3.5:1.5: 1 Mixing, the added ingredients include the following final concentration components: GM-CSF 120ng / mL, glutamine 2.5mmol / L, astragalus polysaccharide 0.3mg / L, notoginseng polysaccharide 0.3mg / L, chicken embryo extract 1.5v / v%, ginger juice 1.5v / v%, kiwi fruit juice 1.5v / v%, IL-420ng / mL, TNF-α1000UI / mL, iron citrate 350mg / L and vitamin C 4mg / L.

[0032] Among them, the astragalus polysaccharide is prepared by the following method: take the waste residue extracted from total astragalus saponins, add water 4 times the weight of the waste residue, boil for 40 minutes, filter to obtain the first filtrate and the first precipitate, and add 3 times the weight to the first precipitate water, boiled for 40min, filtered to...

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Abstract

The invention discloses a dendritic cell culture medium and a culture method. The dendritic cell culture medium comprises a basic culture medium and additive components, wherein the basic culture medium consists of a glucose injection, a normal saline injection, an amino acid injection and a trace element injection; the additive components comprise the following components in final concentrations:100-200ng / mL of a GM-CSF (Granulocyte-Macrophage Colony Stimulating Factor), 2-4mmol / L of glutamine, 0.2-0.5mg / L of astragalus polysaccharide, 0.2-0.5mg / L of panax notoginseng polysaccharides, 1-3v / v% of embryo extraction of chicken, 1-3v / v% of ginger juice, 1-3v / v% of kiwifruit juice, and the like. The culture medium disclosed by the invention is definite in component, free of animal-based proteins, free of potential safety hazards, meanwhile free of substances with high cost, and easy to popularize; moreover, different components in the culture medium take a synergistic effect into play, and when being adopted to culture dendritic cells, the culture medium is rapid in amplification speed and high in activity.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a dendritic cell culture medium and a culture method. Background technique [0002] Somatic cell therapy refers to a treatment method that applies human autologous, allogeneic or heterologous (non-human) somatic cells, and reinfuses (or implants) them into the human body after extracorporeal manipulation. This in vitro operation includes cell passage, expansion, screening, and drugs or other treatments that can change the biological behavior of cells. The somatic cells after in vitro operation can be used for the treatment of diseases, and can also be used for the diagnosis and prevention of diseases. There are many different types of somatic cell therapy, including in vivo reinfusion of in vitro activated mononuclear cells such as lymphokine-activated killer cells, tumor infiltrating lymphocytes, monocytes, macrophages, or in vitro sensitized killer cells. Dome...

Claims

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Application Information

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IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2500/30C12N2500/32C12N2500/34C12N2500/38C12N2500/76C12N2500/80C12N2501/22C12N2501/23C12N2501/25
Inventor 胡小东熊川粤秦连荣熊惠芳
Owner 重庆斯德姆生物技术有限公司
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