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Glycyrrhiza polysaccharide-containing and animal origin-free low protein culture medium

A technology of licorice polysaccharide and culture medium, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problems of unfavorable cell line culture technology research, high price, complex components, etc., and is conducive to separation and purification. , the effect of promoting biosynthesis and improving expression ability

Inactive Publication Date: 2017-05-31
严志海
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the existing serum-free medium, some cannot achieve high-density culture process; although some can support cell growth well, it may lead to the reduction or even loss of the ability of cells to express products; in addition, the formula of commercial serum-free medium is often It is a commercial secret, and the components are complex and expensive, which is not conducive to the research of specific cell line culture technology, nor can it be directly used in the research, development and optimization of cell culture process

Method used

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  • Glycyrrhiza polysaccharide-containing and animal origin-free low protein culture medium
  • Glycyrrhiza polysaccharide-containing and animal origin-free low protein culture medium
  • Glycyrrhiza polysaccharide-containing and animal origin-free low protein culture medium

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Effect test

Embodiment 1

[0028] Embodiment 1 A kind of non-animal source low-protein culture medium containing licorice polysaccharide

[0029] A licorice polysaccharide-free animal-source low-protein medium, consisting of the following components and their concentrations:

[0030] Licorice polysaccharide 85mg / L, glucose 15g / L, soybean hydrolyzate 2.5g / L, putrescine 10.5mg / L, spermine 0.75mg / L, ornithine 0.75mg / L, sodium pyruvate 72mg / L, glutamine Amide 0.45g / L, Alanine 8.5mg / L, Arginine 0.17g / L, Asparagine 9.5mg / L, Aspartic Acid 9.35mg / L, Cysteine ​​33.5mg / L, Cystine Acid 46.87mg / L, glutamic acid 15.5mg / L, glycine 40.25mg / L, histidine 35mg / L, isoleucine 87.5mg / L, leucine 95.5mg / L, lysine 135mg / L , phenylalanine 65mg / L, proline 27.5mg / L, serine 45mg / L, threonine 77.6mg / L, tryptophan 8.3mg / L, tyrosine 78mg / L, valine 96.5mg / L, biotin 0.0075mg / L, vitamin D 0.135mg / L, folic acid 5.5mg / L, ferrous sulfate 3.23mg / L, ferric nitrate 0.45mg / L, EDTA·2Na8.5mg / L, ethanolamine 4.75mg / L L, glycerin 0.45mg / L, cod...

Embodiment 2

[0041] Embodiment 2 A kind of non-animal source low-protein culture medium containing licorice polysaccharide

[0042] A licorice polysaccharide-free animal-source low-protein medium, consisting of the following components and their concentrations:

[0043] Licorice polysaccharide 45mg / L, glucose 20g / L, soybean hydrolyzate 3g / L, putrescine 15mg / L, spermine 1.5mg / L, ornithine 1.35mg / L, sodium pyruvate 75mg / L, glutamine 0.75 g / L, alanine 10mg / L, arginine 0.25g / L, asparagine 12mg / L, aspartic acid 10mg / L, cysteine ​​45mg / L, cystine 55mg / L, gluten Glycine 18mg / L, Glycine 45mg / L, Histidine 40mg / L, Isoleucine 100mg / L, Leucine 150mg / L, Lysine 185mg / L, Phenylalanine 70mg / L, Proline Acid 30mg / L, Serine 50mg / L, Threonine 80mg / L, Tryptophan 15mg / L, Tyrosine 100mg / L, Valine 100mg / L, Biotin 0.01mg / L, Vitamin D 2.50mg / L L, folic acid 8mg / L, ferrous sulfate 4.5mg / L, ferric nitrate 0.5mg / L, EDTA·2Na10mg / L, ethanolamine 8.5mg / L, glycerin 0.45mg / L, cod liver oil 0.2mg / L, cholesterol 0.15mg / L ...

Embodiment 3

[0045] Embodiment 3 A kind of non-animal source low-protein culture medium containing licorice polysaccharide

[0046] A licorice polysaccharide-free animal-source low-protein medium, consisting of the following components and their concentrations:

[0047] Licorice polysaccharide 150mg / L, glucose 10g / L, soybean hydrolyzate 1.5g / L, putrescine 0.5mg / L, spermine 0.1mg / L, ornithine 0.045mg / L, sodium pyruvate 65mg / L, glutamine Amide 0.3g / L, alanine 5mg / L, arginine 0.15g / L, asparagine 7mg / L, aspartic acid 8mg / L, cysteine ​​25mg / L, cystine 30mg / L , glutamic acid 6mg / L, glycine 20mg / L, histidine 25mg / L, isoleucine 50mg / L, leucine 55mg / L, lysine 80mg / L, phenylalanine 50mg / L, Proline 20mg / L, Serine 35mg / L, Threonine 60mg / L, Tryptophan 10mg / L, Tyrosine 60mg / L, Valine 60mg / L, Biotin 0.005mg / L, Vitamin D 0.50 ~2.50mg / L, folic acid 2.50~8mg / L, ferrous sulfate 1.5~4.5mg / L, ferric nitrate 0.1~0.5mg / L, EDTA·2Na 1.5~10mg / L, ethanolamine 3.5~8.5mg / L, glycerin 0.3~0.45mg / L, cod liver oil 0.1~...

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Abstract

The invention belongs to the technical field of cell culture in vitro and particularly relates to a glycyrrhiza polysaccharide-containing and animal origin-free low protein culture medium. The culture medium is prepared from the following components: glycyrrhiza polysaccharide, glucose, soybean hydrolysate, putrescine, spermine, ornithine, sodium pyruvate, amino acid, biotin, vitamin D, folic acid, ferrous sulfate, ferric nitrate, EDTA (Ethylene Diamine Tetraacetic Acid).2Na, cholamine, glycerinum, oleum morrhuae, cholesterol, linoleic acid, thymidine, hypoxanthine, hydrocortisone, sodium selenite, tocopherol, sodium bicarbonate, calcium chloride, magnesium chloride, magnesium sulfate, potassium chloride, sodium dihydrogen phosphate, sodium chloride, disodium hydrogen phosphate, zinc sulfate and copper sulfate. The culture medium disclosed by the invention comprises added glycyrrhiza polysaccharide, and does not contain any added supplementary animal origin protein and / or recombinant animal protein; rapid proliferation of cells can be realized, and the expression ability of the cells is effectively improved.

Description

technical field [0001] The invention belongs to the technical field of in vitro cell culture, and in particular relates to an animal-sourced low-protein culture medium containing licorice polysaccharide. Background technique [0002] As the scale of mammalian cell culture expands and the demand for biopharmaceuticals grows, the development of high-quality cell culture media is becoming increasingly important. Currently, cell culture medium formulations require the addition of a range of additives, including undefined components such as fetal calf serum (FCS), several animal-derived proteins, and / or bovine protein hydrolysates. Cell culture media often require the addition of serum or serum-derived substances such as albumin, transferrin or insulin, which may contain unwanted substances that can contaminate the cell culture and the resulting biological product. In addition, human serum-derived supplements must be tested for all known serum-infectious viruses including hepati...

Claims

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Application Information

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IPC IPC(8): C12N5/078
Inventor 严志海
Owner 严志海
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