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Preparation method of proliferation culture medium for 3D simulation culture of stem cells

A technology for proliferation medium and stem cell proliferation, which is applied in the field of production of proliferation medium for 3D simulation culture, can solve the problems of lack of medium for stem cell 3D simulation culture, unsuitable for exercise physiological process, unclear composition of cell lysate, etc.

Pending Publication Date: 2022-02-18
华子昂
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since 2D culture technology is based on the expansion mode of static culture, the cells obtained from 2D culture cannot adapt to the physiological process of human body movement after entering the human body. The stem cell industry is transitioning to 3D simulation culture technology
Since the medium currently on the market is designed for 2D culture technology based on petri dishes and culture bottles, there is a lack of medium for 3D simulation culture of stem cells
In addition, many stem cell culture media also use cell lysates (or organelle lysates) with unclear components, and very few still use animal serum components, which have a great impact on the prevention of cross-infection of infectious diseases and the study of the mechanism of stem cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025]The formulation of the proliferation medium for 3D simulation culture of stem cells is as follows: its composition includes: inorganic salts, amino acids, sugars, lipids, nucleosides, vitamins, cell metabolites, cell proliferation factors, water, and the osmotic pressure is 300 mOsmol / kg H2O, pH7.4, specific ingredients, inorganic salts: 0.03% potassium ions, 0.5% sodium ions, 0.01% calcium ions, 0.003% magnesium ions, 0.0001% iron ions, 0.16% bicarbonate ions, sulfuric acid Root ion 0.001%, phosphate ion 0.02%, chloride ion 0.4%. Natural amino acids: alanine 0.0005%, arginine 0.0005%, asparagine 0.0005%, aspartic acid 0.0005%, cysteine ​​0.0005%, glutamine 0.03%, glutamic acid 0.0005%, glycine 0.0005% , Histidine 0.0005%, Isoleucine 0.0005%, Leucine 0.0005%, Lysine 0.0005%, Methionine 0.0005%, Phenylalanine 0.0005%, Proline 0.0005%, Serine 0.0005%, Threonine 0.0005%, Tryptophan 0.0005%, Tyrosine 0.0005%, Valine 0.0005%. Sugars: glucose 0.1%, sodium pyruvate 0.02%. L...

Embodiment 2

[0044] The formulation of the proliferation medium for 3D simulation culture of stem cells is as follows: its composition includes: inorganic salts, amino acids, sugars, lipids, nucleosides, vitamins, cell metabolites, cell proliferation factors, water, and the osmotic pressure is 300 mOsmol / kg H2O, pH 7.4, specific ingredients, inorganic salts: 0.01% potassium ions, 0.6% sodium ions, 0.003% calcium ions, 0.0002% magnesium ions, 0.0002% iron ions, 0.1% bicarbonate ions, sulfuric acid Root ion 0.0005%-0.005%, phosphate ion 0.005%-0.05%, chloride ion 0.5%. Natural amino acids: alanine 0.01%, arginine 0.03%, asparagine 0.01%, aspartic acid 0.01%, cysteine ​​0.01%, glutamine 0.06%, glutamic acid 0.01%, glycine 0.01% , Histidine 0.01%, Isoleucine 0.01%, Leucine 0.01%, Lysine 0.01%, Methionine 0.01%, Phenylalanine 0.01%, Proline 0.01%, Serine 0.01%, Threonine 0.01%, Tryptophan 0.01%, Tyrosine 0.01%, Valine 0.01%. Sugars: glucose 0.01%, sodium pyruvate 0.05%. Lipids: Lecithin 0.5...

Embodiment 3

[0063] The formulation of the proliferation medium for 3D simulation culture of stem cells is as follows: its composition includes: inorganic salts, amino acids, sugars, lipids, nucleosides, vitamins, cell metabolites, cell proliferation factors, water, and the osmotic pressure is 300 mOsmol / kg H2O, pH 7.4, specific ingredients, inorganic salts: 0.02% potassium ions, 0.7% sodium ions, 0.004% calcium ions, 0.01% magnesium ions, 0.0002% iron ions, 0.1% carbonate ions, sulfate ions Ion 0.005%, Phosphate ion 0.05%, Chloride ion 0.6%. Natural amino acids: alanine 0.01%, arginine 0.03%, asparagine 0.01%, aspartic acid 0.01%, cysteine ​​0.01%, glutamine 0.06%, glutamic acid 0.01%, glycine 0.01% , Histidine 0.01%, Isoleucine 0.01%, Leucine 0.01%, Lysine 0.01%, Methionine 0.01%, Phenylalanine 0.01%, Proline 0.01%, Serine 0.01%, Threonine 0.01%, Tryptophan 0.01%, Tyrosine 0.01%, Valine 0.01%. Sugars: glucose 0.01%, sodium pyruvate 0.05%. Lipids: Lecithin 0.3%, Cholesterol 0.05%, Ste...

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PUM

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Abstract

The invention provides a proliferation culture medium for 3D simulation culture of stem cells, the proliferation culture medium is free of serum addition, definite in composition and suitable for 3D simulation perfusion culture of stem cells, and comprises inorganic salt, amino acid, saccharides, lipids, nucleosides, vitamins, cell metabolites, cell proliferation factors and water, the osmotic pressure of a culture solution is 270-330mOsmol / kg.H2O, and the pH value of the culture solution is 7.2-7.4.

Description

technical field [0001] The invention belongs to the field of medicine, in particular to a method for making a proliferation medium for 3D simulation culture. Background technique [0002] Embryonic stem cells are a kind of primitive undifferentiated cells with unlimited proliferation ability and unlimited differentiation potential. They are the core driving force of contemporary medical progress and medical efficiency improvement. Mesenchymal stem cells and adult stem cells are currently the most popular stem cell research directions. They have the advantages of easy acquisition, easy scale-up and culture, strong differentiation ability, and immune regulation. Therefore, they have made great progress in regenerative medicine and stem cell therapy and have shown great potential. market application potential. [0003] The 2D culture technology based on petri dishes and culture bottles has made great contributions to stem cell culture, completed the technical transition of ste...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N2500/90C12N2500/14C12N2500/12C12N2500/16C12N2500/24C12N2500/32C12N2500/34C12N2500/30C12N2500/36C12N2500/40C12N2500/38C12N2500/35C12N2501/11C12N2501/135C12N2513/00
Inventor 华子昂刘宝全朱美瑛万君兴竹添
Owner 华子昂
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