Psoriasis skin repairing agent prepared by mixing stem cell extract and traditional Chinese medicine extract and application thereof
A skin repairing agent and extract technology, applied in the field of psoriasis skin repairing agent, to achieve the effect of less fibroblasts, inhibiting growth and increasing blood vessel density
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Embodiment 1
[0036] The preparation of embodiment 1 skin restoration agent of the present invention
[0037] Step 1: Preparation of umbilical cord mesenchymal stem cell extract
[0038] (1) Take out the umbilical cord tissue of the newborn, remove blood vessels, epithelial membranes and blood clots, separate Wharton's block, and cut it into 1mm 3 After size, put it into a 10cm sterile petri dish; digest with DMEM medium containing 0.1% collagenase II; after filtration, use DMEM medium containing 10% fetal bovine serum FBS for subculture, and replace it with serum-free medium on the third day Subculture in DMEM medium, once every 2 days; take a bottle of cells from passage 3 for subsequent treatment;
[0039] (2) digest the cells with DMEM medium containing 0.05M EDTA, and wash with PBS solution;
[0040] (3) The cells were resuspended in serum-free DMEM medium, and lysed and broken by ultrasonic (120W, 15S*20 times, 15S interval), the broken product was centrifuged at 12000rpm for 15 min...
Embodiment 2
[0049] The preparation of embodiment 2 skin restoration agent of the present invention
[0050] 1. Experimental method
[0051] (1) Primary culture of umbilical cord mesenchymal stem cells;
[0052] (2) Umbilical cord mesenchymal stem cells were mixed with 2*10 5 cells / ml into a 12-well plate for experiments, and adherent culture for at least 24 hours;
[0053] (3) Add the skin repair agent prepared in Example 1 of the present invention (the mass concentration is respectively 0%, 0.5%, 1%, 2%, 4%, 10% and 15%), and add the cell damage agent bleomycin as comparison;
[0054] (4) Perform flow cytometry detection on the cells to detect cell apoptosis and cell viability (CCK8 method).
[0055] 2. Experimental results
[0056] For the CCK8 experiment of mesenchymal stem cells, the cytotoxicity of the skin repair agent of the present invention at a concentration of 4% is 6.0%, and the cytotoxicity at a concentration of 10% is 15.2%, indicating that its cytotoxicity is small, an...
Embodiment 3
[0057] Example 3 The cell experiment that the skin repair agent of the present invention inhibits the growth of fibroblasts and promotes the growth of epidermal stem cells and vascular endothelial cells
[0058] 1. Experimental method
[0059] (1) Primary culture of different cells, including fibroblasts, epidermal stem cells and vascular endothelial cells;
[0060] (2) Divide cells into 2*10 5 Spread into 12-well plates for experiments, 4 wells for each type of cells, and culture on the wall for at least 24 hours;
[0061] (3) Add the skin repair agent (mass concentration 4%) of the present invention to each kind of cell respectively, and add bleomycin (final concentration is 2 μ g / ml) as the cell damage agent, the concrete adding method is:
[0062]
[0063] (4) Perform flow cytometry detection on all cells to detect cell apoptosis and cell viability (CCK8 method).
[0064] 2. Experimental results
[0065] The results of cell apoptosis in each group are shown in Table...
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