Mesenchymal stem cell ossification osteogenic 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 low specificity of osteogenic induction and differentiation, unsatisfactory induction efficiency, etc., and achieve improved osteogenic induction and differentiation efficiency, high-efficiency osteogenic induction and differentiation, and preparation method simple effect

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

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Problems solved by technology

[0006] In order to solve the problems in the prior art that the induction efficiency is not ideal and the specificity of osteogenic induction and differentiation is not strong, the inventor screened through a large number of tests and found an unexpected discovery: by adding a certain concentration of resveratrol and

Method used

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  • Mesenchymal stem cell ossification osteogenic differentiation culture medium and preparation method thereof
  • Mesenchymal stem cell ossification osteogenic differentiation culture medium and preparation method thereof
  • Mesenchymal stem cell ossification osteogenic differentiation culture medium and preparation method thereof

Examples

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Embodiment 1

[0029] Example 1 Formula Screening of Mesenchymal Stem Cell Osteogenic Induction and Differentiation Medium

[0030] The inventors conducted a large number of test screenings on the mesenchymal stem cell osteogenic differentiation medium. Take the 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 the induction medium 1 and induction medium were added respectively. Medium 3, induction medium 4 and induction medium 5 were changed every 3 days. After induction and culture for 4 weeks, the expression level of osteoblast-related gene OPN was identified by qPCR, and the results were as follows: figure 1 shown.

[0031] from figure 1 It can be seen that adding a certain concentration of resveratrol and puerarin to the osteogenic differentiation medium of mesenchymal ...

Embodiment 2

[0032] Example 2 Mesenchymal Stem Cell Osteogenic Induction Differentiation Medium

[0033] Mesenchymal stem cell osteogenic 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 , ascorbic acid 150 μM, β-glycerol phosphate 10 mM, dexamethasone 10 nM, resveratrol 10 μM and puerarin 0.1 μM.

[0034] Preparation method: Add FBS, glutamine, penicillin-streptomycin, ascorbic acid mother solution, β-glycerophosphate mother solution, dexamethasone mother solution, resveratrol mother solution and puerarin mother solution to the DMEM / F12 medium according to the above concentrations , stir evenly, pass through the membrane to sterilize.

[0035] Preparation of ascorbic acid mother solution: take 10 g of ascorbic acid, dissolve it in DMEM / F12 medium, prepare 15 mM mother solution, and store at -20°C. Preparation of β-glycerol phosphate mother ...

Embodiment 3

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

[0037]Mesenchymal stem cell osteogenic differentiation medium, including DMEM / F12 medium, also includes the following components and their concentrations: FBS 15% by volume, glutamine 1.5% by volume, penicillin-streptomycin 1% by volume , ascorbic acid 200 μM, β-glycerol phosphate 20 mM, dexamethasone 10 nM, resveratrol 15 μM and puerarin 0.05 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 ossification osteogenic differentiation culture medium, and belongs to the technical field of stem cells. The mesenchymal stem cell ossification osteogenic 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%, 100-1000 [mu]M ascorbic acid, 5-50 mM glycerol phosphate, 5-50 nM dexamethasone, 5-50 [mu]M resveratrol, and 0.05-0.5 [mu]M puerarin. The mesenchymal stem cell ossification osteogenic 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 osteogenic differentiation medium and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are important members of the stem cell family, derived from the mesoderm and ectoderm in the early stages of development, and belong to pluripotent stem cells. MSCs were originally discovered in the bone marrow, and have received extensive attention because of their multi-directional differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and self-replication. Under specific induction conditions in vivo or in vitro, mesenchymal stem cells can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelial and other tissue cells. It has multi-directional differentiation potential and can be used as an ideal seed cell for the repair of...

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马岩岩
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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