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A kind of stem cell serum-free medium containing small molecules of traditional Chinese medicine and preparation method thereof

A serum-free medium and small molecule technology, applied in the field of cell culture, can solve problems such as improper collection and means of FBS, harm to human safety, heterologous serum residue, etc., and achieve the protection of stem cells, no difference between batches, and clear ingredients Effect

Active Publication Date: 2022-05-27
EXPERIMENTAL RES CENT CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as a heterologous substance, FBS has many shortcomings and hidden dangers, and is not suitable for the cultivation of human-derived mesenchymal stem cells: first, FBS is a mixture, which contains beneficial nutrients needed to maintain cell growth, and may also contain viruses, toxins, etc. , these harmful substances may be carried into the human body with the addition of FBS; secondly, FBS also has the risk of being contaminated by mad cow disease; thirdly, a large number of FBS collections and improper methods may cause potential animal ethics issues; finally, the residue of heterologous serum, in cells After transplantation, it may trigger a severe immune response, endangering human safety
In addition, the use of double antibodies may have toxic and side effects on sensitive primary extracted cells

Method used

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  • A kind of stem cell serum-free medium containing small molecules of traditional Chinese medicine and preparation method thereof
  • A kind of stem cell serum-free medium containing small molecules of traditional Chinese medicine and preparation method thereof
  • A kind of stem cell serum-free medium containing small molecules of traditional Chinese medicine and preparation method thereof

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Embodiment 1

[0023] The present embodiment provides a method for preparing a stem cell serum-free medium containing small molecules of traditional Chinese medicine, comprising the following steps:

[0024] (1) Mix all the raw materials to obtain a concentrate, and at -20 o Store under the condition of C until use; the concentration of each raw material is: recombinant human insulin 5mg / L, vitamin C 20mg / L, human transferrin 7mg / L, human serum albumin 1g / L, recombinant human fibronectin 15mg / L , human epidermal growth factor EGF 5μg / L, human basic fibroblast growth factor bFGF 5μg / L, platelet-derived growth factor PDGF-BB 5μg / L, breviscapine 10nM, ginsenoside Rg3 50nM, baicalin 0.1mg / L , Salvianolic acid B10nM, Berberine 20nM;

[0025] (2) Take and save at 0 o In the DMEM high-glucose medium under the condition of C, add the concentrated solution obtained in step (1) according to the weight ratio of 1:1000 (that is, add 1 mL of concentrated solution to 1 L of medium, the same below), stir...

Embodiment 2

[0028] The present embodiment provides a method for preparing a stem cell serum-free medium containing small molecules of traditional Chinese medicine, comprising the following steps:

[0029] (1) Mix all the raw materials to obtain a concentrate, and at -20 o Store under the condition of C until use; the concentration of each raw material is: recombinant human insulin 10mg / L, vitamin C 50mg / L, human transferrin 15mg / L, human serum albumin 1g / L, recombinant human fibronectin 20mg / L , Human epidermal growth factor EGF 10μg / L, human basic fibroblast growth factor bFGF 10μg / L, platelet-derived growth factor PDGF-BB 20μg / L, breviscapine 200nM, ginsenoside Rg3 100nM, baicalin 0.4mg / L , Salvianolic acid B200nM, Berberine 100nM;

[0030] (2) Take and save in 4 o In the DMEM high-glucose medium under the condition of C, the concentrated solution obtained in step (1) is added according to the weight ratio of 1:1000, and the mixture is uniformly stirred to obtain the raw material of t...

Embodiment 3

[0033] The present embodiment provides a method for preparing a stem cell serum-free medium containing small molecules of traditional Chinese medicine, comprising the following steps:

[0034] (1) Mix all the raw materials to obtain a concentrate, and at -20 o The concentration of each raw material is: recombinant human insulin 15mg / L, vitamin C 100mg / L, human transferrin 15mg / L, human serum albumin 5g / L, recombinant human fibronectin 30mg / L , human epidermal growth factor EGF 20μg / L, human basic fibroblast growth factor bFGF 20μg / L, platelet-derived growth factor PDGF-BB 20μg / L, breviscapine 500nM, ginsenoside Rg3 500nM, baicalin 0.5mg / L , Salvianolic acid B400nM, Berberine 500nM;

[0035] (2) Take and save in 8 o In the DMEM high glucose medium under the condition of C, the concentrated solution obtained in step (1) is added according to the weight ratio of 1:1000, and the mixture is uniformly stirred to obtain the raw material of the pre-culture medium;

[0036] (3) ster...

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Abstract

The invention relates to a stem cell serum-free culture medium containing traditional Chinese medicine small molecules and a preparation method thereof. The medium includes the following raw materials: recombinant human insulin, vitamin C, recombinant human transferrin, human serum albumin, recombinant human fibronectin, human epidermal growth factor EGF, human basic fibroblast growth factor bFGF, platelet-derived growth factor Factors PDGF‑BB, scutellarin, ginsenoside Rg3, baicalin, salvianolic acid B, and berberine. The culture medium includes nutrient factors required by cells and human growth-promoting small molecules without heterologous proteins, and can provide a good environment for maintaining cell growth and proliferation. By adding scutellarin, ginsenoside Rg3, baicalin, salvianolic acid B and berberine, five small molecules of traditional Chinese medicine, it can significantly improve the adhesion of mesenchymal stem cells, promote cell proliferation rate, maintain cell surface phenotype and The cells have multiple differentiation potentials and can better protect stem cells.

Description

technical field [0001] The invention belongs to the field of cell culture, and in particular relates to a stem cell serum-free medium containing traditional Chinese medicine small molecules and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are considered as potential biological materials for regenerative medicine. MSCs have the advantages of rapid propagation, multi-lineage differentiation under induced conditions, and no ethical issues. In recent years, whether at home or abroad, the application of MSCs to transplant human body to treat major diseases has been approved for clinical trials, such as ischemic stroke, rheumatoid arthritis, multiple sclerosis, idiopathic pulmonary fibrosis. chemotherapy and graft-versus-host disease. [0003] The standardized production process and culture of cells are the basis of MSCs research. The medium is a key factor affecting the growth and proliferation of MSCs. The conventional stem cell c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775
Inventor 陈鹏苏晗杨洪军邓世文崔钊黎彩凤
Owner EXPERIMENTAL RES CENT CHINA ACAD OF CHINESE MEDICAL SCI
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