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Mammalian cell growth medium and additive thereof

A mammalian and culture medium technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of pathogen infection, non-specificity, cell contamination, etc.

Inactive Publication Date: 2016-11-23
YANTAI CELLZONE BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since RPMI-1640 medium often adds serum when culturing animal cells, it is easy to cause cell contamination or pathogen infection
At the same time, because the RPMI-1640 medium does not have the specificity of the cultured cell types, it will cause a relative decrease in the cell proliferation ability for some special cells.

Method used

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  • Mammalian cell growth medium and additive thereof
  • Mammalian cell growth medium and additive thereof
  • Mammalian cell growth medium and additive thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1: From the cultivation of mononuclear cells (PBMC) in human peripheral blood

[0052] (1-1) Preparation of velvet active polysaccharide

[0053] Antler active polysaccharide additive (A) was prepared by dissolving 2 mg of velvet active polysaccharide in 1 mL of PBS.

[0054] (1-2) Proliferation of mononuclear cells (PBMC) in human peripheral blood

[0055] Containing amino acids (all of the above amino acids), inorganic salts (all of the above inorganic salts), vitamins (all of the above vitamins) and other additives (adenosine 5' monophosphate, corticosterone, ethanolamine, D-galactose , D-glucose, insulin, reduced glutathione, lipoic acid, hypoxanthine, phenol red, progesterone, putrescine, pyruvate, thymidine, triiodothyronine, transferrin, selenite Add 7×10 -4 % 2-mercaptoethanol (2-Me), 100 U / mL penicillin·streptomycin (prepared in PBS) to obtain a serum-free medium (B1).

[0056] (1-3) The A prepared in (1-1) was added to the serum-free medium at vari...

Embodiment 2

[0072] Embodiment 2: from the cultivation of human adipose tissue-derived stem cells (ADMSCs)

[0073] (2-1) Preparation of velvet active polysaccharide

[0074] Antler active polysaccharide additive (A) was prepared by dissolving 2 mg of velvet active polysaccharide in 1 mL of PBS.

[0075] (2-2) Proliferation of human adipose tissue-derived stem cells (ADMSCs)

[0076] Remove hypoxanthine and thymidine and a part of inorganic salts (copper sulfate, iron nitrate (III), iron sulfate, magnesium chloride, disodium hydrogen phosphate, zinc sulfate) from the basal medium prepared in Example 1, in the obtained Add 100 U / mL penicillin·streptomycin (prepared in PBS) to the basal medium to obtain serum-free medium (B2).

[0077] (2-3) The A prepared in (2-1) was added to the serum-free medium at various concentrations to obtain a serum-free medium (C2).

[0078]Specifically, A was added at the following concentrations.

[0079] Add 0.01mM A serum-free medium in medium B2;

[0080...

Embodiment 3

[0093] Example 3: Culture from Human Bone Marrow Stem Cells (MSCs)

[0094] (3-1) Preparation of velvet active polysaccharide

[0095] Antler active polysaccharide additive (A) was prepared by dissolving 2 mg of velvet active polysaccharide in 1 mL of PBS.

[0096] (3-2) Proliferation of human bone marrow stem cells (MSCs)

[0097] Remove transforming growth factor, granulocyte colony-stimulating factor, epidermal growth factor and triiodothyronine and a part of inorganic salts (copper sulfate, iron nitrate (III), iron sulfate , magnesium chloride, disodium hydrogen phosphate, zinc sulfate), and 100 U / mL penicillin·streptomycin (prepared in PBS) was added to the obtained basal medium to obtain a serum-free medium (B3).

[0098] (3-3) A prepared in (2-1) was added to the serum-free medium at various concentrations to obtain a serum-free medium (C3).

[0099] Specifically, A was added at the following concentrations.

[0100] Add 0.01mM A serum-free medium in medium B3;

[...

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Abstract

The present invention relates to a medium used for culturing mammalian cells in vitro and additives for constituting the medium, which can effectively make mammalian somatic cells without using serum when culturing mammalian somatic cells proliferation. By mixing velvet antler active polysaccharides in the medium, mammalian somatic cells can be efficiently proliferated even when the medium is completely free of serum.

Description

technical field [0001] The invention belongs to the field of cell culture medium, and in particular relates to a culture medium used for culturing mammalian cells in vitro and additives for constituting the culture medium. Background technique [0002] Mammalian cells are the basis of research in the fields of cell therapy and regenerative medicine. The cells include cells with multi-differentiation functions of mammals including humans and cells differentiated from these cells. Advancing the cultivation of these cells so that they proliferate rapidly is key to the research and productivity of this field. [0003] It is known that after mammalian cells are isolated, they are usually cultured and proliferated in a medium supplemented with about 10% serum. However, as the serum, it usually comes from autologous blood of cells to be cultured (for example, patient's blood from which somatic cells to be cultured) or serum from other sources such as bovine serum. However, when ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/078C12N5/0775C12N5/074
Inventor 翟红利李晓宇张红梅张淑敏杨小平
Owner YANTAI CELLZONE BIOTECH CO LTD
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