A kind of medical dressing of stem cell extract and preparation method thereof
A stem cell and extract technology, applied in the field of biomedicine, can solve the problems of breeding bacteria, hindering the rapid healing of wounds, slow absorption, etc., to achieve the effect of promoting skin tissue, excellent synergy, and weak immunogenicity
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Embodiment 1
[0064] Embodiment 1, preparation stem cell extract
[0065] (1) Preparation of BMSCs extract
[0066] Take the purchased embryo-derived BMSCs cell P4 out of the liquid nitrogen refrigerator, quickly place it in a 37°C water bath and shake it to melt, centrifuge the cryovial at 1500rpm for 5min, discard the supernatant, and resuspend with 1ml recovery medium ;
[0067] The recovery medium is DMEM-F12 medium containing 15% fetal bovine serum, and the cell count is 8*10 5 Cells were seeded on 75cm 2 In the culture bottle, add the medium in the culture bottle to 15ml;
[0068] Place the flask at 37°C, 5% CO 2 Cultivate in an incubator, change the medium after 24 hours (DMEM-F12 medium containing 10% fetal bovine serum), and then change the medium every 2 days. After 3-5 days, the cell fusion reaches more than 90% for passage;
[0069] When passaging, first discard the medium, wash twice with 5-10ml PBS, add 2ml 0.05% trypsin for digestion, shake gently to make the trypsin cov...
Embodiment 2
[0088] Embodiment 2, preparation collagen freeze-dried powder
[0089] (1) Extraction of fish skin collagen;
[0090] Step 1, tissue crushing: After thawing the skin of adult salamander (artificial breeding) weighing 2-3 catties, add water so that the weight ratio of fish skin to water is 10:1, crush and grind the mixture, and then crush Ultrasonic action is added while grinding, the ultrasonic intensity is controlled at 750 watts / liter, and the ultrasonic treatment time is 40min to obtain a slurry;
[0091] Step 2, remove salt-soluble impurities: mix the slurry with sodium chloride solution (mass concentration 5%), the weight ratio of slurry and sodium chloride solution is 1:5, stir well, soak for 15h, 3000rpm, centrifuge for 20min, Remove the supernatant. get the first precipitation;
[0092] Step 3, remove acid-soluble impurities: mix the first precipitate with citric acid solution (mass concentration is 10%), the weight ratio of the first precipitate and acid solution is...
Embodiment 3
[0102] Embodiment 3, preparation medical dressing
[0103] 100g of medical dressings contains: bone marrow mesenchymal stem cell (BMSCs) extract lyophilized powder prepared in Example 1 (mass fraction 0.2%), collagen lyophilized powder prepared in Example 2 (mass fraction 0.1%); sodium alginate (mass fraction 2%); concentrated vitamin C powder (10%); normal saline (mass fraction 87.7%).
[0104] When in use, just mix the powder and water evenly. 10g of the dressing contains 20mg of BMSCs extract freeze-dried powder, 10mg of collagen freeze-dried powder, 0.2g of sodium alginate, 1g of concentrated vitamin C powder, and 8.77g of normal saline.
[0105] 10g dressing can be applied no larger than 10 x 10cm 2 wound surface.
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