Collagen composition and collagen film patch for improving allergic symptoms
A collagen and composition technology, applied in the field of biomedicine, 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 are separated and cultured and then subcultured. The culture medium used for primary culture and subculture is DMEM / F12 medium + 10% volume fraction of fetal bovine serum. When the primary When the confluence of the cells reaches 80%, discard the culture medium, then wash the cells with PBS phosphate buffered saline for 3 times, then add 0.04% EDTA+0.25% trypsin digestion solution, the amount of digestion solution added is 2mL / 10cm 2 The area of the bottom of the culture dish, after 80% of the cells become round, add the culture solution to stop the digestion, the volume of the culture solution is 5 times the volume of the digestion solution. Centrifuge the digested liquid at 600g for 5 minutes, remove the supernatant, add culture medium to resuspend the cells, and adjust the cell density to 1×10 5 / mL, the cell suspe...
Embodiment 2
[0064] The preparation of embodiment 2 collagen composition
[0065] Take P3-P8 generation cells, discard the culture medium and wash with PBS 3 times, according to 2mL / 10cm 2 Add 0.04% EDTA+0.25% trypsin digestion solution to digest the cells according to the ratio of the bottom area of the culture dish, and then use an ultrasonic breaker adjusted to 9000 cycles to sonicate the digested cell suspension for 8 minutes.
[0066] After the treatment, the suspension was centrifuged with a centrifugal force of 19,000 g for 25 minutes, the supernatant was discarded, the precipitate was resuspended by adding PBS containing 500 mmol Nacl, and the resuspension was filtered through a filter membrane with a pore size of 0.45 μm.
[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 sterilized by filtration ...
Embodiment 3
[0071] The preparation of embodiment 3 collagen composition
[0072] Take P3-P8 generation cells, discard the culture medium and wash with PBS 3 times, according to 2mL / 10cm 2 Add 0.04% EDTA+0.25% trypsin digestion solution to digest the cells according to the ratio of the bottom area of the culture dish, and then use an ultrasonic breaker adjusted to 9000 cycles to sonicate the digested cell suspension for 8 minutes.
[0073] After the treatment, the suspension was centrifuged with a centrifugal force of 15,000 g for 25 minutes, the supernatant was discarded, the precipitate was resuspended by adding PBS containing 700 mmol Nacl, and the resuspension was filtered through a filter membrane with a pore size of 0.5 μm.
[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 w...
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Abstract
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