Method for evaluating or screening hair growth-regulating agent
A technology for hair growth regulators and growth promoters, which is applied in the field of evaluation or screening of hair growth regulators, and can solve the problems of unsuitable mass screening, difficulty in obtaining human hair follicles, unsuitable screening, etc.
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Embodiment 1
[0052] (1) Separation of hair follicles
[0053] As the pig skin, use the skin obtained from a white-haired hybrid pig raised in Japan as a meat pig, cut it to an appropriate size, cut off excess fat, and put it in chlorhexidine under a sterile environment. solution (diluted 5% chlorhexidine solution (manufactured by Sumitomo Pharmaceutical Co., Ltd.) with water to 5 to 20 times), soaked for 5 to 10 minutes, disinfected, and then washed several times with D-PBS.
[0054] Next, under a solid microscope, hair follicles were separated from the treated pig skin with tweezers and a scalpel (FEATHER No 10), and concentrated in the culture medium. As the culture medium, RPMI1640 medium (Gibco cat. No. 11835-030), William's Medium E medium (Gibco cat. No. 1255 1-032) and DMEM / HamF12 (1:1) medium (Gibco cat. No. 11039-021).
[0055] (2) Effects of DHT on pig hair follicle organ culture
[0056] Pig hair follicles were obtained from the pig's back, flanks, abdomen, groin, buttocks, a...
Embodiment 2
[0068] Evaluation of embodiment 2 hair growth regulator
[0069] (1) t-flavanone and TGF-β2
[0070] Pig buttock hair follicles were obtained in the same manner as in Example 1. As a medium, RPMI1640 (Gibco cat. No. 11835-030) medium was used. A medium in which a penicillin / streptomycin (Gibco cat. No. 15140-122) solution was added at a ratio of 1% to RPMI1640 medium was used.
[0071] Using the above-mentioned RPMI1640 medium, t-flavanone (trans-3) which was confirmed to have a hair growth-promoting effect in organ culture of beard hair follicles of rats as an active ingredient of a hair growth-promoting agent not belonging to pharmaceuticals was prepared. , 4′-dimethyl-3-hydroxyflavanone (Hotta et al., Fragrance Journal, Vol. 31 No. 2, 33-40 (2003))), the final concentration in the culture medium 1 μM (for the synthesis method, refer to Japanese Patent Laid-Open No. 2000-198779). Furthermore, TGF-β2 (R&D System, Inc., model number: 102-B2-001 / CF), which was confirmed to ...
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