Collagen composition for improving allergy symptoms and collagen patch
A technology of collagen and composition, which is applied in the field of biomedicine and can solve problems such as human harm
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Embodiment 1
[0062] Example 1 Preparation of mesenchymal stem cell extract
[0063] The primary human umbilical cord or umbilical cord blood-derived mesenchymal stem cells were isolated and cultured and then subcultured. The culture medium used for primary culture and subculture was DMEM / F12 medium + 10% fetal bovine serum. When the cell confluence reaches 80%, discard the culture medium, then wash the cells with PBS phosphate buffer three times, then add 0.04% EDTA + 0.25% trypsin digestion solution, the amount of digestion solution is 2mL / 10cm 2 The bottom area of the petri dish, after 80% of the cells become round, add the culture solution to terminate the digestion, and the added volume of the culture solution is 5 times the added volume of the digestion solution. Centrifuge the digested liquid at 600g for 5 minutes, remove the supernatant, add the culture medium to resuspend the cells, and adjust the cell density to 1×10 5 / mL, the cell suspension was added to a new dish, recorded ...
Embodiment 2
[0064] Example 2 Preparation of collagen composition
[0065] Take the P3~P8 generation cells, discard the culture medium and wash 3 times with PBS, according to 2mL / 10cm 2 0.04% EDTA+0.25% trypsin digestion solution was added to the ratio of the bottom area of the petri dish to digest the cells, and then the digested cell suspension was sonicated at 9,000 cycles with a sonicator for 8 minutes.
[0066] After the treatment, the suspension was centrifuged at 19000 g for 25 minutes, the supernatant was discarded, PBS containing 500 mmol NaCl was added to resuspend the precipitate, and the resuspended liquid was filtered through a 0.45 μm pore size filter.
[0067] The filtrate was collected and concentrated by ultrafiltration using a filter membrane with a molecular weight cut-off of 5KD, and the concentrated volume was 1 / 5 of the original volume.
[0068] The concentrated solution was filtered and sterilized with a 0.22um μm pore size filter, and then the protein concentrati...
Embodiment 3
[0071] Example 3 Preparation of collagen composition
[0072] Take the P3~P8 generation cells, discard the culture medium and wash 3 times with PBS, according to 2mL / 10cm 2 0.04% EDTA+0.25% trypsin digestion solution was added to the ratio of the bottom area of the petri dish to digest the cells, and then the digested cell suspension was sonicated at 9,000 cycles with a sonicator for 8 minutes.
[0073] After the treatment, the suspension was centrifuged at 15000 g for 25 minutes, the supernatant was discarded, PBS containing 700 mmol NaCl was added to resuspend the precipitate, and the resuspended liquid was filtered through a 0.5 μm pore size filter.
[0074] The filtrate was collected and concentrated by ultrafiltration using a filter membrane with a molecular weight cut-off of 5KD, and the concentrated volume was 1 / 10 of the original volume.
[0075]The concentrated solution was sterilized by filtration with a 0.22um μm pore size filter, and then the protein concentrati...
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