Cosmetic composition containing galactomyces-derived exosome as active ingredient
A cosmetic composition, galactose technology, applied in the direction of medical preparations, cosmetics, cosmetic preparations containing active ingredients, etc., can solve problems such as foreign body sensation, skin problems, skin redness or dermatitis
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Embodiment 1
[0044] Example 1: Preparation of galactosomal exosomes
[0045] Commercially available galactosaccharomyces strains G. candidum and G. geotrichum used for brewing or fermentation were cultured with shaking, and the conditioned medium was subsequently pelleted by centrifugation and precipitated galactose was removed yeast cells.
[0046] The supernatant of the galactomyces conditioned medium obtained by the above procedure was filtered through a 0.22-μm filter to remove impurities such as cell debris, waste, and large particles. Galactomyces-derived exosomes were isolated from filtered Galactomyces conditioned medium by a tangential flow filtration (TFF) method.
[0047] The size of isolated Galactomyces-derived exosomes was analyzed by transmission electron microscopy (TEM). As shown in Figures 1A and 1B, it was determined that the isolated Galactomyces-derived exosomes were nano-sized vesicles. The size and concentration of Galactomyces-derived exosomes were analyzed by na...
Embodiment 2
[0048] Example 2: Evaluation of the delivery ability of Galactomyces-derived exosomes to dermal fibroblasts
[0049] In order to examine whether galactomyces-derived exosomes would be delivered to human dermal fibroblasts (purchased from CEFO Co., Ltd.), the following analysis was performed. To fluorescently stain the membranes of the galactomyces-derived exosomes prepared in Example 1, the exosomes were reacted with PKH67 fluorescent dye (purchased from Sigma-Aldrich). After the reaction, the reaction solution was fractionated with a MW3000 column (purchased from ThermoFisher Scientific) to remove unstained free PHK67 in the exosome membrane. A negative control was prepared by reacting PKH67 fluorescent dye with buffer solution and fractionating the reaction product with a MW3000 column. Exosomes stained with PKH67 were incubated with pre-cultured human dermal fibroblasts, and fluorescence microscopy was then used to observe whether exosomes would be delivered into the cells...
Embodiment 3
[0050] Example 3: Evaluation of the effect of stimulating collagen production
[0051] Human dermal fibroblasts (purchased from CEFO Co., Ltd.) dispersed in DMEM medium containing fetal bovine serum were dispensed into multiwell plates, and then cultured for 24 hours, and cultured in serum-free medium for another 24 hours. Hour. Thereafter, the galactomyces-derived exosomes prepared in Example 1 were diluted in different concentrations of serum-free medium, and then human dermal fibroblasts were treated with each dilution and cultured. Thereafter, the medium was collected and centrifuged, and then the centrifuged medium was prepared. The amount of collagen synthesized by human dermal fibroblasts and accumulated in the medium was measured using an EIA kit for type I procollagen C-peptide (PIP) (purchased from Takara Bio). The total protein amount of cell lysates was measured with BCA protein assay kit (purchased from ThermoFisher Scientific). Normalize the amount of collagen...
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