Human mesenchymal stem cell cartilage-formation induced differentiation culture medium and preparation method
A technology of mesenchymal stem cells and induced differentiation, applied in the field of culture medium, can solve the problems of chondrogenic induction efficiency and specificity, long induction time, etc., and achieve the effects of high degree of induced differentiation, simple preparation method, and convenient use
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Embodiment 1
[0043] Example 1: Formula screening of human mesenchymal stem cell chondrogenic differentiation medium
[0044] The inventors of the present invention screened the components of the human mesenchymal stem cell ru-induced differentiation medium through a large number of experiments. Human mesenchymal stem cells were cultured and expanded in low-sugar DMEM medium containing 10% FBS. Trypsinize the cells, resuspend the cells with the chondrogenic differentiation medium described in the patent claims, the concentration is 2X10 7 cells / mL; add 10 μL cell suspension dropwise to a six-well plate, 5 drops per well; place the cells in the incubator for 2 hours, and add differentiation-inducing medium 1, differentiation-inducing medium 2, and differentiation-inducing medium to each well. Medium 3, and induction differentiation medium 4, the medium was changed every 3 days. At 2 and 3 weeks of induction, the expression levels of chondrocyte-related genes CollagenII and Aggrecan were de...
Embodiment 2
[0045] Example 2: Preparation of human mesenchymal stem cell chondrogenic differentiation medium
[0046] Human mesenchymal stem cell chondrogenic differentiation medium, including HG-MEM medium, and the following components and their concentrations:
[0047] Fetal bovine serum (FBS)
10% (volume percentage)
sodium pyruvate
100mg / ml
50μg / ml
20μg / ml
100nM
ITS
1% (volume percentage)
TGF-β 3
5ng / ml
Insulin Growth Factor-2 (IGF-2)
20nM
[0048] Preparation method: Add FBS, sodium pyruvate mother solution, ascorbic acid mother solution, proline mother solution, dexamethasone mother solution, ITS mother solution, TGF-β to HG-MEM medium according to the above concentrations 3 Mix the mother liquor and the IGF-2 mother liquor evenly, and filter and sterilize with a 0.22 μm filter membrane.
[0049] Preparation of ascorbic acid mother solution: Take 50 mg...
Embodiment 3
[0050] Example 3: Human mesenchymal stem cell chondrogenic differentiation medium
[0051] Human mesenchymal stem cell chondrogenic differentiation medium, including HG-MEM medium, and the following components and their concentrations:
[0052]
[0053]
[0054] Preparation method: similar to the preparation method provided in Example 2.
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