A Lactobacillus plantarum that improves sensitive facial skin and repairs the skin barrier
A technology of Lactobacillus plantarum and viable count, applied in the field of microorganisms, can solve problems such as side effects, complex drug action mechanism, dependence, etc., and achieve the effects of improving survival rate, improving barrier damage, and promoting cell migration.
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[0060] 2. Preparation of Topical Probiotic Samples
[0061] (1) Probiotic fermentation supernatant: take the cultured probiotics by centrifugation at 8000r for 15 min to take the supernatant; adjust the pH to 7.2-7.4; filter and sterilize with a 0.22 μm disposable filter, and store in a -20°C refrigerator after packaging.
[0062] (2) Probiotic intracellular substance: take 10 mL of cultured probiotic bacteria to centrifuge to collect bacterial mud, add 1 mL of PBS solution to dissolve; add liquid nitrogen to grind and crush, collect bacterial mud suspension; add 1 mL of PBS solution to dissolve, ultrasonically crushed; centrifuge at 8000 r The supernatant was collected for 10 min; sterilized by filtration with a 0.22 μm disposable filter, and stored in a refrigerator at -20° C. after packaging (0.1 g of wet bacteria mud obtained 1 mL of intracellular supernatant).
[0063] Complete medium: 5% fetal bovine serum (FBS), 100 U / mL penicillin, 100 mg / mL streptomycin, 95% DMEM medi...
Embodiment 1
[0064] Example 1: Screening and identification of Lactobacillus plantarum
[0065] Specific steps are as follows:
[0066] (1) Isolation and screening of Lactobacillus plantarum
[0067] (1) collecting kimchi samples from different regions, and enriching the samples in MRS medium containing sorbitol for 12h;
[0068] (2) The enriched sample in step (1) is subjected to gradient dilution and then coated on the MRS solid plate added with 0.02% olfactory cresyl violet, and cultured for 24-48 hours;
[0069] (3) select the single colony with obvious discoloration circle on the MRS plate in step (2) and conform to the basic form of lactic acid bacteria to carry out streak purification on the MRS solid plate, and screen and isolate the lactic acid bacteria;
[0070] (4) The single colony on the plate in step (3) is picked and cultured in the liquid MRS medium for 24 hours, and then Gram-stained, and Gram-positive bacteria are selected for the subsequent test.
[0071] (2) Prelimin...
Embodiment 2
[0092] Example 2: Effect of Lactobacillus plantarum on SDS-induced HaCaT cell survival
[0093] Specific steps are as follows:
[0094] After rewarming the cryopreserved HaCaT cells at 37°C, centrifuge at 1000 rpm for 5 min to take the pellet; after washing the pellet once with complete medium, resuspend the cells in complete medium and count to obtain a cell resuspend; Inoculated into a 10cm petri dish at 37°C, gas phase containing 5% (v / v) CO 2 The cells were cultured in a cell incubator, and the complete medium was replaced the next day and continued at 37°C with 5% (v / v) CO in the gas phase. 2 cultured in a cell incubator. Cells were passaged when they had grown to 70% to 80% density of the dish.
[0095] HaCaT cells with good growth status were selected, the HaCaT cells were digested with trypsin at a concentration of 2.5 g / L, centrifuged, resuspended in complete medium, and counted to obtain a cell resuspension; the cell resuspension was 5× 10 3 Inoculate each well ...
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