Lactobacillus plantarum capable of improving facial sensitive skin and repairing skin barriers
A technology of Lactobacillus plantarum and microbial strains, applied in the field of microorganisms, can solve problems such as dependence, complex drug action mechanism, side effects, etc.
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[0060] 2. Preparation of topical probiotic samples
[0061] (1) Probiotic fermentation supernatant: take the cultured probiotics and centrifuge at 8000rr for 15 minutes to get the supernatant; adjust the pH to 7.2-7.4; filter and sterilize with a 0.22 μm disposable filter, store in a -20°C refrigerator after aliquoting.
[0062] (2) Intracellular content of probiotics: Take 10mL of cultured probiotics and collect the bacteria sludge by centrifugation, add 1mL PBS solution to dissolve; add liquid nitrogen to grind and crush, collect the bacteria sludge suspension; add 1mL PBS solution to dissolve, ultrasonically crush; centrifuge at 8000r The supernatant was collected for 10 minutes; sterilized by filtration with a 0.22 μm disposable filter, and stored in a -20°C refrigerator after aliquoting (0.1 g of wet sludge obtained 1 mL of intracellular supernatant).
[0063] Complete medium: 5% fetal bovine serum (FBS), 100 U / mL penicillin, 100 mg / mL streptomycin, 95% DMEM medium.
Embodiment 1
[0064] Example 1: Screening and Identification of Plantarum Lactobacillus
[0065] Specific steps are as follows:
[0066] (1) Isolation and screening of Lactobacillus plantarum
[0067] (1) Collect samples of kimchi produced in different regions, and enrich the samples in MRS medium containing sorbitol for 12h;
[0068] (2) Gradiently dilute the enriched sample in step (1) and spread it on the MRS solid plate added with 0.02% cresyl violet, and incubate for 24-48 hours;
[0069] (3) Select a single colony with a clear color circle on the MRS plate in step (2) and conform to the basic form of lactic acid bacteria to the MRS solid plate for streaking and purification, and screen and isolate the lactic acid bacteria;
[0070] (4) Pick a single colony on the plate in step (3) and culture it in liquid MRS culture solution for 24 hours, then carry out Gram staining, and select Gram-positive bacteria for subsequent tests.
[0071] (2) Preliminary identification of Lactobacillus p...
Embodiment 2
[0092] Example 2: Effect of plantarum lactobacillus on the survival of HaCaT cells induced by SDS
[0093] Specific steps are as follows:
[0094] After rewarming the frozen HaCaT cells at 37°C, centrifuge at 1000rpm for 5min, and take the pellet; wash the pellet once with complete medium, resuspend the cells with complete medium and count to obtain a cell suspension; Inoculate into a 10cm Petri dish at 37°C with 5% (v / v) CO in the gas phase 2 cultured in a cell incubator, and the next day, the complete medium was replaced and continued at 37°C with 5% (v / v) CO in the gas phase. 2 cultured in a cell culture incubator. Cells were subcultured when they reached 70%-80% of the density of the culture dish.
[0095] Select HaCaT cells in a good growth state, digest the HaCaT cells with trypsin at a concentration of 2.5g / L, centrifuge, resuspend with complete medium, and perform cell counting to obtain a cell suspension; 10 3 Each well was inoculated in a 96-well culture plate, ...
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