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Human mesenchymal stem cell adipogenesis inducing and differentiating culture medium and preparation method thereof

A technology for mesenchymal stem cells and induction of differentiation, applied in the field of culture medium, can solve the problems of unsatisfactory adipogenic induction efficiency and specificity, long induction time, etc., and achieve the effect of shortening induction time, simple preparation method, and high-efficiency adipogenic induction and differentiation

Inactive Publication Date: 2018-09-28
安徽瑞杰赛尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems in the prior art that the efficiency and specificity of adipogenic induction are not ideal and the induction time is long, the present invention provides a culture method capable of efficiently and stably inducing the directed adipogenic differentiation of a variety of human tissue-derived mesenchymal stem cells in vitro. base and its preparation method

Method used

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  • Human mesenchymal stem cell adipogenesis inducing and differentiating culture medium and preparation method thereof
  • Human mesenchymal stem cell adipogenesis inducing and differentiating culture medium and preparation method thereof
  • Human mesenchymal stem cell adipogenesis inducing and differentiating culture medium and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Example 1: Formula screening of human mesenchymal stem cell adipogenic differentiation medium

[0045] The inventors of the present invention screened the components of the culture medium for adipogenic induction and differentiation of human mesenchymal stem cells through a large number of experiments. Human mesenchymal stem cells were cultured and expanded in low-sugar DMEM medium containing 10% FBS. Digest the cells with trypsin, take the cells and inoculate them into 6-well plates according to 2×104 cells / cm2, and grow them in growth medium (HG-DMEM medium + 10% FBS (volume percentage) + 1% glutamine (volume percentage)). When the basic confluence is reached, replace the cost-patented adipogenic induction culture medium for induction, and change the medium once every 3 days, so that the culture cycle lasts until 2 weeks (14 days). At the 1st and 2nd week of induction, use realtime PCR to detect the adipogenic cells. Gene C / EBPβ and PPARγ2 expression levels. The res...

Embodiment 2

[0046] Example 2: Preparation of Human Mesenchymal Stem Cell Adipogenic Induction Differentiation Medium

[0047] Human mesenchymal stem cell adipogenic differentiation medium, including HG-MEM medium, and the following components and their concentrations:

[0048] Fetal bovine serum (FBS)

10% (volume percentage)

Glutamine

1% (volume percentage)

Indomethacin

200μM

insulin

1μg / ml

1-methyl-3-isobutylxanthine

100μM

Dexamethasone

100nM

spermine

1μM

[0049] Preparation method: Add FBS, glutamine mother solution, indomethacin mother solution, 1-methyl-3-isobutylxanthine mother solution, dexamethasone mother solution, spermine to the HG-DMEM medium according to the stated concentration Mix the mother liquor evenly, and filter it through a 0.22 μm membrane to obtain the product.

[0050] Preparation of indomethacin mother solution: take 10 mg of indomethacin, dissolve it in DMSO, prepare 80 mM mother...

Embodiment 3

[0051] Example 3: Human mesenchymal stem cell adipogenic differentiation medium

[0052] Human mesenchymal stem cell adipogenic differentiation medium, including HG-MEM medium, and the following components and their concentrations:

[0053] Fetal bovine serum (FBS)

[0054] Preparation method: similar to the preparation method provided in Example 2.

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Abstract

The invention discloses a human mesenchymal stem cell adipogenesis inducing and differentiating culture medium and a preparation method thereof. The human mesenchymal stem cell adipogenesis inducing and differentiating culture medium is produced by the following components of an alpha-MEM / HG-DMEM culture medium, 5-50% of percent by volume of fetal calf serum, 0.5-10% of percent by volume of glutamine, 100-400 [mu]M volume of indomethacin, insulin with the concentration of 0.1-20 [mu]g / ml, 10-200 [mu]M volume of 1-methyl-3-isobutyl xanthine, 10-200 nM volume of dexamethasone and 0.1-20 [mu]M volume of spermine. The preparation method includes the following steps that a culture dish is cleaned; the culture medium is provided, and the alpha-MEM / HG-DMEM culture medium is prepared in the culture dish according to a formula; materials are mixed; and mixed liquor is filtered. Multiple histologic origin human mesenchymal stem cells including human mesenchymal stem cells, umbilical cord mesenchymal stem cells and adipose tissue-derived stromal cells are induced to the directional differentiation of adipogenesis cells; differentiating and inducing time of the human mesenchymal stem cells isshortened, the preparation method is convenient, and the differentiating and the inducing of human mesenchymal stem cell adipogenesis can be achieved stably and efficiently.

Description

technical field [0001] The invention relates to the technical field of culture medium, in particular to a human mesenchymal stem cell adipogenic differentiation culture medium and a preparation method thereof. Background technique [0002] There are mesenchymal stem cells (MSCs) in various tissues of the human body, such as bone marrow, umbilical cord, and fat. Fat, bone, cartilage, muscle, liver, nerve, skin and other types of tissue cells have the potential to differentiate. In recent years, studies have also found that this type of mesenchymal stem cells participates in the functional conditions of the body's immune system and promotes tissue damage repair. Based on the above characteristics, a large number of studies have begun to pay attention to the application potential of mesenchymal stem cell therapy in various tissue regeneration and organ function damage repair. Among them, plastic surgery application research based on the directional adipogenic differentiation c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2500/30C12N2500/32C12N2500/40C12N2500/46C12N2501/30C12N2501/33C12N2506/1353C12N2506/1369C12N2506/1384
Inventor 罗乐朱灏
Owner 安徽瑞杰赛尔生物科技有限公司
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