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Mesenchymal stem cell adipogenic differentiation culture medium and preparation method thereof

A technology for mesenchymal stem cells and induced differentiation, applied in the field of stem cells, can solve the problems of unsatisfactory induction efficiency and achieve the effects of improved adipogenic induction and differentiation efficiency, high adipogenic induction and differentiation, and simple preparation methods

Inactive Publication Date: 2015-08-12
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that the induction efficiency in the prior art is not ideal enough, the inventor screened through a large number of experiments, and unexpectedly found that by adding a certain concentration of Fasudil hydrochloride, a new adipogenic effect of mesenchymal stem cells was obtained. Induction differentiation medium, the induction efficiency of this medium is greatly improved, and the specificity of adipogenic differentiation is improved, and a large number of adipogenic cells can be obtained through the induction of this medium

Method used

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  • Mesenchymal stem cell adipogenic differentiation culture medium and preparation method thereof
  • Mesenchymal stem cell adipogenic differentiation culture medium and preparation method thereof
  • Mesenchymal stem cell adipogenic differentiation culture medium and preparation method thereof

Examples

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Embodiment 1

[0028] Example 1 Formula screening of mesenchymal stem cell adipogenic differentiation medium

[0029] The inventors conducted experimental screening on the adipogenic induction differentiation medium of mesenchymal stem cells. Take P3 generation bone marrow mesenchymal stem cells at 5000 cells / cm 2 The density of the cells was inoculated in a six-well plate, and cultured with DMEM / F12 medium containing 10% (v / v) FBS. When the confluence of the cells reached 80%, the medium was discarded, and induction medium 1 and induction medium 1 were added respectively. Medium 3, the medium was changed every 3 days. After induction and culture for 3 weeks, the expression level of the adipogenic cell-related gene FABP4 was identified by qPCR, and the results were as follows: figure 1 shown.

[0030] from figure 1 It can be seen that adding a certain concentration of Fasudil hydrochloride to the adipogenic induction and differentiation medium of mesenchymal stem cells in the prior art...

Embodiment 2

[0032] Example 2 Mesenchymal stem cell adipogenic induction differentiation medium

[0033] Mesenchymal stem cell adipogenic differentiation medium, including DMEM / F12 medium, also includes the following components and their concentrations: FBS 10% by volume, glutamine 1% by volume, penicillin-streptomycin 1% by volume , indomethacin 200 μM, insulin 100 nM, dexamethasone 10 nM, 3-isobutyl-1-methylxanthine 1 μM and fasudil hydrochloride 0.1 μM.

[0034] Preparation method: Add FBS, glutamine, antibiotics, indomethacin mother solution, insulin mother solution, dexamethasone mother solution, 3-isobutyl-1-methylxanthine to the DMEM / F12 medium according to the stated concentration The mother liquor and the fasudil hydrochloride mother liquor are stirred evenly, and the bacteria are passed through a membrane to remove bacteria.

[0035]Preparation of indomethacin mother solution: take 100mg, dissolve in 50% methanol, make 20mM mother solution, and store at -20°C. Preparation of ...

Embodiment 3

[0036] Example 3 Mesenchymal Stem Cell Adipogenic Induction Differentiation Medium

[0037] Mesenchymal stem cell adipogenic differentiation medium, including DMEM / F12 medium, also includes the following components and their concentrations: FBS 15% by volume, glutamine 1% by volume, antibiotics 1% by volume, indomethacin Dixime 150 μM, insulin 50 nM, dexamethasone 10 nM, 3-isobutyl-1-methylxanthine 2 μM and fasudil hydrochloride 0.2 μM.

[0038] Preparation method: similar to the preparation method in Example 2.

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Abstract

The present invention provides a mesenchymal stem cell adipogenic differentiation culture medium, and belongs to the technical field of stem cells. The mesenchymal stem cell adipogenic differentiation culture medium comprises a DMEM / F12 culture medium, and further comprises FBS with a volume percentage of 5-50%, glutamine with a volume percentage of 0.5-5%, antibiotic with a volume percentage of 0.5-5%, 50-500 [mu]M indometacin, 50-500 nM insulin, 5-50 nM dexamethasone, 0.5-5 [mu]M 3-isobutyl-1-methylxanthine, and 0.05-0.5 [mu]M fasudil hydrochloride. The mesenchymal stem cell adipogenic differentiation culture medium of the present invention has advantages of high inducing efficiency and the like.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and relates to a mesenchymal stem cell adipogenic differentiation medium and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of adult stem cells with obvious plasticity and multi-directional differentiation potential. Under the regulation of different growth factors, they can differentiate into different tissue cell types derived from the three germ layers, such as fat cells, osteoblasts, liver-like cells, cardiomyocyte-like cells, glial cells, neuron cells and islet-like cells, etc. In addition, human mesenchymal stem cells do not express major histocompatibility class II antigens, but express a small amount of major histocompatibility class I antigens, have low immunogenicity, have little transplant rejection, and have immunoregulatory effects. Human amniotic membrane after transplantation The nutritional factors secreted by mesenchymal s...

Claims

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Application Information

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IPC IPC(8): C12N5/077
Inventor 陈海佳王一飞葛啸虎戴国胜卢瑞珊
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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