Medicinal composition for inducing stem cells to differentiate into cartilage and application of medicinal composition
A stem cell differentiation and composition technology, applied in the field of medicine, can solve problems such as high price, and achieve the effects of promoting proliferation, inducing and promoting differentiation into chondrocytes, and having strong pharmacological effects
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Embodiment 1
[0036] Embodiment 1 is prepared medicinal liquid
[0037] 1. Prepare puerarin solution
[0038] Accurately weigh 4.36mg puerarin, dissolve in 1mL DMSO, and prepare 10-2 M puerarin mother liquor, the mother liquor was subpackaged and stored in a refrigerator at -20°C, filtered through a disposable sterile syringe filter with a pore size of 0.22 μm before use, and diluted with deionized water as a solvent in sequence to a concentration of 10 μM, 1 μM, Five different concentrations of puerarin, 0.1 μM, 0.01 μM, and 0.001 μM, were administered to the solution.
[0039] 2. Preparation of tetrandrine solution
[0040] Accurately weigh 6.28mg tetrandrine, dissolve in 1mL DMSO, and prepare 10 -2 M powder tetrandrine mother liquor, divide the mother liquor, store in -20°C refrigerator, filter it through a 0.22μm pore size disposable sterile syringe filter before use, and use deionized water as a solvent to dilute it successively to a concentration of 10μM and 5μM , 2.5μM, 1.25μM, 0....
Embodiment 2
[0050] Example 2 Isolation, culture and identification of SD rat bone marrow mesenchymal stem cells
[0051] 1. Experimental animals and reagents
[0052] Healthy SPF-grade Sprague Dawley (SD) male rats, 4-6 weeks old, were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. (permit number: SCXK (Beijing) 2016-0006).
[0053] Certified Foetal Bovine Serum Qualtified for human MSC (fetal bovine serum for stem cell culture grade), Bioilgical Industries, the United States; MEM-ALPHA medium, Bioilgical Industries, the United States; DMEM high-sugar, low-sugar phenol red-free medium, Sigma-Aldrich Company, the United States ; 0.25% trypsin digest (containing 2.21mM EDTA), Corning, the United States; CD29 (PE Hamster Anti-Rat CD29), CD45 (PE-Cy TM 5 Mouse Anti-Rat CD45), CD54 (PE Mouse Anti-Rat CD54), CD90 (APC Mouse Anti-Rat CD90 / MouseCD90.1), CD11b (FITC Mouse Anti-Rat CD11b), BD Biosciences, the United States; RecombinantHuman BMP -2. Recombinant Human...
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