A serum-free medium for human adipose-derived mesenchymal stem cells and preparation method thereof

A serum-free culture medium and stem cell technology, applied in the field of stem cells, can solve the problems of poor cell adhesion, unsatisfactory cell proliferation rate, complex components, etc., and achieve simple preparation methods, good cell proliferation rate and cell stemness, cost reduction effect

Active Publication Date: 2019-03-29
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 problems of poor cell adhesion, complex components, and unsatisfactory cell proliferation rate in the prior art, the inventor obtained a serum-free medium for human adipose-derived mesenchymal stem cells through a large number of tests and screening. The use of culture bottles does not need to be coated, the cell adhesion is good, the cell proliferation rate is high, and the naive morphology and characteristics of stem cells are maintained, which reduces the cost

Method used

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  • A serum-free medium for human adipose-derived mesenchymal stem cells and preparation method thereof
  • A serum-free medium for human adipose-derived mesenchymal stem cells and preparation method thereof
  • A serum-free medium for human adipose-derived mesenchymal stem cells and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Effects of medium containing different concentrations of PGG on cell proliferation

[0030] 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.

[0031] Dissolve PGG powder with DMSO into 100mM stock solution, and dilute PGG stock solution with high-glucose DMEM basal medium to 20μM, 15μM, 10μM, 7.5μM, 5μM, 2.5μM, 1.25μM, 0.63μM, 0.31μM, 0.16μM, 0.08 μM and 0.04 μM PGG solutions.

Embodiment 2

[0033] Example 2 Serum-free medium of human adipose-derived mesenchymal stem cells

[0034] 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 4mM, reduced glutathione 12μg / mL, recombinant human insulin 8 μg / mL, human serum albumin 1 mg / mL, transferrin 3.5 μg / mL and 1,2,3,4,6-O-pentagalloyl glucose 10 μM.

[0035] Preparation method: Add L-glutamine, glutathione, recombinant human insulin, human serum albumin, transferrin and 1,2,3,4,6-O - Pentagalloylglucose, mixed evenly, passed through a membrane to remove bacteria.

Embodiment 3

[0036] Example 3 Serum-free medium of human adipose-derived mesenchymal stem cells

[0037] 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 2mM, reduced glutathione 10μg / mL, recombinant human insulin 5 μg / mL, human serum albumin 0.7 mg / mL, transferrin 5 μg / mL and 1,2,3,4,6-O-pentagalloyl glucose 5 μM.

[0038] The preparation method is similar to Example 2.

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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, glutathione, recombinant human insulin, human serum albumin, transferrin 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 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 Patents(China)
IPC IPC(8): C12N5/0775
Inventor 王一飞廖晓凤王巧利刘秋英
Owner 广东美赛尔细胞生物科技有限公司
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