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A serum-free medium for mesenchymal stem cells containing berberine

A serum-free medium and quality stem cell technology, applied in the field of stem cells, can solve the problems of unsatisfactory cell proliferation effect, prolonged cell growth cycle, and high cell proliferation rate, and achieves delayed entry into the death phase, good cell proliferation rate, and simple preparation method. Effect

Active Publication Date: 2019-02-01
广东美赛尔细胞生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] In order to solve the defects of poor cell adhesion, short cell growth cycle, and unsatisfactory cell proliferation effect in the prior art, the inventor obtained a serum-free medium for mesenchymal stem cells containing berberine through a large number of tests and screenings. The use of this medium does not require pre-coating of culture vessels, and the cells have good adhesion and high cell proliferation rate, which can delay MSCs from entering the decay phase, prolong the growth cycle of cells, and reduce costs

Method used

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  • A serum-free medium for mesenchymal stem cells containing berberine
  • A serum-free medium for mesenchymal stem cells containing berberine
  • A serum-free medium for mesenchymal stem cells containing berberine

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Embodiment 1

[0027] Embodiment 1 Preparation of culture medium of the present invention

[0028] The inventors conducted a large number of tests to screen and obtain the optimal formula of the serum-free medium for mesenchymal stem cells containing berberine provided by the present invention. The preparation of culture medium of the present invention comprises the following steps:

[0029](1) Preparation of basal medium: Take 1 small bag of low-sugar DMEM dry powder, dissolve it in 800ml ultrapure water, and stir to obtain low-sugar DMEM basal medium;

[0030] (2) Add the following components to the basal medium: 40 mg / L berberine, 10 mg / ml basic fibroblast growth factor, 12 mg / ml epidermal growth factor, 10 μg / mL glutathione, recombinant Human insulin 2mg / ml, human serum albumin 10mg / ml, transferrin 6μg / mL, fibronectin 10mg / L, putrescine 5mg / L, aminoethanol 1mg / L, hydrocortisone 30μg / L, pyruvate Sodium 0.07mg / ml and sodium bicarbonate 3.7g / L, mix well, stir to dissolve;

[0031] (3) Ad...

Embodiment 2

[0032] Embodiment 2 Preparation of culture medium of the present invention

[0033] The preparation method of the medium in Example 2 of the present invention is the same as that in Example 1, specifically adding the following components to the basal medium: berberine 5mg / L, basic fibroblast growth factor 3mg / ml, epidermal growth factor 3mg / ml, glutathione 1μg / mL, recombinant human insulin 1mg / ml, human serum albumin 5mg / ml, transferrin 1μg / mL, fibronectin 5mg / L, putrescine 5mg / L, aminoethanol 0.37 mg / L, hydrocortisone 10μg / L, sodium pyruvate 0.02mg / ml and sodium bicarbonate 3.7g / L.

Embodiment 3

[0034] Embodiment 3 Preparation of culture medium of the present invention

[0035] The preparation method of the medium in Example 3 of the present invention is the same as that in Example 1, specifically adding the following concentrations of components to the basal medium: berberine 110mg / L, basic fibroblast growth factor 20mg / ml, epidermal growth factor 20mg / ml, glutathione 10μg / mL, recombinant human insulin 10mg / ml, human serum albumin 15mg / ml, transferrin 10μg / mL, fibronectin 15mg / L, putrescine 30mg / L, aminoethanol 4mg / L, hydrocortisone 50μg / L, sodium pyruvate 0.2mg / ml and sodium bicarbonate 3.7g / L.

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Abstract

The invention provides berberine-containing serum-free medium for mesenchymal stem cells, comprising a DMEM basic medium and further comprising, by concentration, 5-110mg / L berberine, 3-20mg / ml basic fibroblast growth factors, 3-20mg / ml epidermal growth factors, 1-10Mug / mL glutathione, 1-10mg / ml recombinant human insulin, 5-15mg / ml human serum albumin, 1-10Mug / mL transferrin, 5-15mg / L fibronectin, 5-30mg / L putrescine, 0.37-4mg / L aminoethanol, 10-50Mug / mL hydrocortisone, and 0.02-0.2mg / ml sodium pyruvate. The invention belongs to the technical field of stem cells; the medium has good cell adherence, high cell proliferation rate and capable of delaying decline phase of MSCs (mesenchymal stem cells) and prolong growth period of the cells.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and relates to a serum-free medium for mesenchymal stem cells containing berberine. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells belonging to the mesoderm, which have strong proliferation ability and multi-lineage differentiation potential, and can not only differentiate into osteoblasts, cartilage cells, and Cells, fat cells, and also have the ability to differentiate into muscle cells, liver cells, hematopoietic cells, nerve cells, islet cells and other cells. MSC has a wide range of sources, is easy to isolate, culture, expand and purify, has low immunogenicity, and has no restrictions on moral and ethical issues, so it has great application prospects in tissue engineering, gene therapy and immunotherapy. [0003] The traditional stem cell medium containing animal serum has brought many disadvantages to the production and scientific resea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775
Inventor 王一飞梁峰王巧利
Owner 广东美赛尔细胞生物科技有限公司
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