Application of chick embryo bioactive peptide in preparing product for protecting cell activity
A chicken embryo active peptide and cell technology, which is applied in the application field of the product to achieve the effects of improving fine lines, active protection and removing red blood.
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Embodiment 1
[0054] (1) Hatching chicken embryos: Take SPF grade fertilized eggs and put them into the incubator for incubation. The incubation temperature is controlled at 37.5°C, the relative humidity is 60%, and the eggs are automatically turned every 2 hours. After hatching to 16 days, take out the fertilized eggs , put it in a freezer at -20°C, stop the incubation and save it for later use.
[0055] (2) Homogenization and cell crushing: surface disinfection of the hatched eggs, removal of egg shells, taking out the chicken embryos, homogenizing the chicken embryos with a tissue homogenizer at 4°C, pouring them into a sterilized sealed box, Put it in a freezer at -30°C for 72 hours, and after thawing, perform cell ultrasonic disruption in an environment of 4°C, add an equal amount of water for injection, and place it in a freezer at 4°C for use.
[0056] (3) Enzymatic hydrolysis and centrifugation: Take the refrigerated solution after cell crushing, heat it in a water bath to 50°C, adj...
Embodiment 2
[0067] Cell viability was determined by MTT colorimetry. The detection principle is that succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple crystalline formazan (Formazan) and deposit in the cells, while dead cells do not have this function. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and its light absorption value is measured at a wavelength of 550nm by an enzyme-linked immunosorbent detector, which can indirectly reflect the number of living cells. Within a certain range of cell numbers, the amount of MTT crystal formation is proportional to the number of living cells.
[0068] MTT assay for cell viability:
[0069] Human fibroblasts (HDF cells) (the sixth generation) were treated with 1×10 4 Individuals / well were inoculated in 96-well plates;
[0070] The experiment was divided into experimental group and blank control group;
[0071] After 24 hours, the experimental group was given different...
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