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Serum-free medium for human adipose-derived stem cells and preparation method thereof

A serum-free medium and stem cell technology, applied in the field of stem cells, can solve the problems of poor cell adherence and unsatisfactory cell proliferation rate, achieve simple preparation method, good cell proliferation rate and cell stemness, and promote rapid adherence Effect

Active Publication Date: 2015-12-02
广东美赛尔细胞生物科技有限公司
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Problems solved by technology

[0009] In order to solve the problems of poor cell adhesion and unsatisfactory cell proliferation rate in the prior art, the inventor obtained a new serum-free medium for human adipose-derived mesenchymal stem cells through a large number of tests and screening. The use of this medium No need for culture flasks for coating, good cell adhesion, high cell proliferation rate, good stem cell immature morphology and stem cell characteristics, reducing costs

Method used

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  • Serum-free medium for human adipose-derived stem cells and preparation method thereof
  • Serum-free medium for human adipose-derived stem cells and preparation method thereof
  • Serum-free medium for human adipose-derived stem cells and preparation method thereof

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Embodiment 1

[0030] Embodiment one contains the influence of the culture medium of different concentration PGG on cell proliferation

[0031] The inventor conducted a large number of tests to screen and obtain the serum-free medium formulation of human adipose-derived mesenchymal stem cells provided by the present invention, and further investigated the influence of the concentration of PGG. Dissolve PGG powder with DMSO into 100 mM stock solution, and dilute PGG stock solution into 20 μM, 15 μM, 12.5 μM, 10 μM, 7.5 μM, 5 μM, 2.5 μM, 1.25 μM, 0.63 μM and 0.31 μM with high-glucose DMEM basal medium, respectively. PGG solution.

[0032] The P1 generation human adipose-derived mesenchymal stem cells were treated with 10 4 The density of each / mL was inoculated in a 96-well plate, and the serum-free medium of human adipose-derived mesenchymal stem cells containing different concentrations of PGG (the difference from the serum-free medium of human adipose-derived mesenchymal stem cells provid...

Embodiment 2

[0033] Example 2 Effect of culture medium containing different concentrations of stem cell growth factors on cell proliferation

[0034] The P1 generation human adipose-derived mesenchymal stem cells were treated with 10 4 The density of each / mL was inoculated in a 96-well plate, and the serum-free medium of human adipose-derived mesenchymal stem cells containing different concentrations of stem cell growth factors (serum-free medium of human adipose-derived mesenchymal stem cells provided in Example 6) were respectively used The difference lies in the concentration of stem cell growth factor) culture, 100 μL per well, and the serum-free medium of human adipose-derived mesenchymal stem cells without stem cell growth factor as the control group. Placed at 37°C, 5% CO 2 After 24 hours, use the MTT detection method to measure the absorbance value of each well, compare the absorbance value of the treatment group with stem cell growth factor with the absorbance value of the cont...

Embodiment 3

[0035] Example 3 Serum-free medium of human adipose-derived mesenchymal stem cells

[0036] Serum-free medium for human adipose-derived mesenchymal stem cells, including high-glucose DMEM basal medium, and the following components and their concentrations: L-glutamine 5mM, insulin-like growth factor-110ng / mL, epidermal growth factor 10ng / mL, transforming growth factor-beta 5ng / mL, platelet-derived growth factor 5ng / mL, basic fibroblast growth factor 10ng / mL, transferrin 5μg / mL, vitamin C25μg / mL, human serum albumin 1mg / mL and 1,2,3,4,6-O-pentagalloyl glucose 10 μM.

[0037] Preparation method: Add L-glutamine, insulin-like growth factor, epidermal growth factor, transforming growth factor, platelet-derived growth factor, fibroblast growth factor, transferrin, Mix vitamin C, human serum albumin and 1,2,3,4,6-O-pentagalloyl glucose, and pass through a membrane to remove bacteria.

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Abstract

The invention provides a serum-free medium for human adipose-derived stem cells, comprising a DMEM basic medium and further comprising L-glutamine, insulin-like growth factors, epidermal growth factors, transforming growth factors, platelet-derived growth factors, fibroblast growth factors, transferrin, vitamin C, human serum albumin and 1, 2, 3, 4, 6-O-pentagalloylglucose. The invention belongs to the technical field of stem cells; the serum-free medium for human adipose-derived stem cells provided by the invention is capable of promoting quick adherence and proliferation of the human adipose-derived stem cells and keeping good stem cell pedomorphism and stem cell characteristics.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and relates to a serum-free culture medium of human adipose mesenchymal stem cells and a preparation method thereof. Background technique [0002] Human adipose-derived stem cells (ADSCs) are a kind of stem cells isolated from adipose tissue with multipotential differentiation potential. Studies have found that ADSCs cells can proliferate stably in vitro, and at the same time have the advantages of abundant sources, large reserves in the body, easier extraction than bone marrow, a large number of stem cells can be obtained from a small amount of tissue, and a low death rate. Mesenchymal stem cells derived from adult adipose tissue have high clinical application value and are widely used in the fields of tissue engineering and regenerative medicine. [0003] 1,2,3,4,6-O-pentagalloylglucose (1,2,3,4,6-Pentagalloylglucose, PGG) is a compound extracted from the medicinal plant Phyllanthus emblica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 王一飞程江王巧利廖晓凤
Owner 广东美赛尔细胞生物科技有限公司
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