Composition for preventing hair loss or promoting hair growth, containing humanin
A technology of hair growth and composition, applied in the direction of medical preparations containing active ingredients, hair care, application, etc., can solve problems such as skin irritation, lack of curative effect and human body stability
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Embodiment 1
[0049] Embodiment 1: preparation and method of material
[0050] 1-1: Prepare Brain Protector
[0051] Humanin (HNG) produced by PEPTIDE INSTITUTE, INC (Minoh, Osaka, Japan) was used in this experiment. The Humanin (HNG) is the Humanin (HNG) in which the 14th amino acid base of the original Humanin (HN) is replaced by glycine (glycine) from serine (serine). Human brain (HNG) was dissolved in sterile triple distilled water to prepare a 0.1 mM human brain (HNG) solution, which was diluted and used.
[0052] 1-2: Culture of dermal papilla cells
[0053] Immortalized dermal papilla cells (Rat vibrissa immortalized dermal papilla cell: Filsell W, et al., 1994) isolated from the whiskers of white mice were treated with 100 units / ml of penicillin-100 μg / ml of streptomycin (Gibco Inc, New York, U.S.) and 10% heat-inactivated fetal bovine serum (heat-inactivated fetal bovine serum) (FBS; Gibco Inc, New York, U.S.) DMEM (Hyclone Inc, U.S.) medium, at 37 ℃ The culture was carried out...
Embodiment 2
[0056] Example 2: Confirmation of the growth and proliferation efficacy of dermal papilla cells by tetrazolium salt (MTT) analysis
[0057] Proliferation of dermal papilla cells was measured using a tetrazolium salt colorimetric assay (MTT assay). The dermal papilla cells of 0.5×104cell / ml were mixed in DMEM (Hyclone Inc, USA) containing 0.5% fetal bovine serum (FBS) and cultured in a 96-well plate (wellplate) for 24 hours, then treated with HNG to make it final Concentrations up to 50nM, 100nM, 200nM and 500nM. Minoxidil sulfate as a positive control group was treated at a concentration of 1 μM. After culturing for 72 hours, 50 μl of 2 mg / ml MTT solution (Sigma, MO, USA) was treated, and the reaction was carried out for 4 hours. After centrifugation after 4 hours, the supernatant was removed, and 200 μl of DMSO was added to dissolve the precipitate. The absorbance of the above-mentioned lysate was measured at 540 nm using VERSAmaxmicroplate reader (Molecular Devices, CA, U...
Embodiment 3
[0060] Example 3: Confirm that Humanin induces hair follicle growth
[0061] Cut the left and right mystacial pads of rats (rat) born 3 weeks old, and put them into E / P buffer (1:1 vol. / vol.solution of Earle's balanced saltssolution and phosphate-buffered saline supplemented with100U penicillin / mL and 100μg streptomycin / ml). The excised mystacial pads were observed with a dissecting microscope and vibrissa follicles were isolated using a scalpel. Put the isolated vibrissae follicles into E medium (William E medium supplemented with 2mM L-glutamine, 10μg / mlinsulin, 50nM hydrocortisone, 100U / ml penicillin and 100μg / ml streptomycin) in 50nM, 100nM Human brain (HNG) was treated and cultured at a concentration of 200nM. New E medium (William E medium) was replaced every 3 days, and Humanin (HNG) was also treated at concentrations of 50 nM, 100 nM and 200 nM.
[0062] Vibrissa follicles in culture were photographed every 7 days using a microscope (Olympus, Japan), and the length ...
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