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5 protocols using follicle dermal papilla cell growth medium kit

1

Human Hair Follicle Dermal Papilla Cell Culture

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Human hair follicle dermal papilla (HDP) cells were purchased from PromoCell (Germany). The passage 3-7 cells were maintained in 5% CO 2 at 37 o C in a follicle dermal papilla cell growth medium kit (PromoCell) and subcultured when the cells reached 70 to 80% confluency. For experiments, the cells were grown in Dulbecco's Modified Eagle's Medium (DMEM; Thermo Fisher Scientific, USA) supplemented with 5% (v/v) fetal bovine serum (FBS; Sigma-Aldrich, USA). The human HaCaT keratinocytes (Thermo Fisher Scientific) were cultured in Epilife (Invitrogen, USA) supplemented with Human Keratinocytes Growth Supplement (HKGS; Invitrogen). 293T cells (American Type Culture Collection, USA) were cultured in DMEM supplemented with 10% (v/v) FBS. TCF/LEF luciferase reporter plasmids were obtained from Promega (USA). LY294002 was purchased from Merck (Germany).
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2

Cultivation of Hair Follicle Dermal Papilla and Endothelial Cells

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HFDPCs were purchased from PromoCell (Heidelberg, Germany) and were cultured in a follicle dermal papilla cell growth medium kit (Promocell, Heidelberg, Germany) supplemented with 1% (v/v) penicillin/streptomycin (P4333, Sigma-Aldrich, St. Louis, MO, USA). HFDPCs were used within four passages. HUVECs were purchased from Thermo Fisher Scientific (USA) and were cultured in endothelial cell basal medium-2 (EBM-2, Lonza, Walkersville, MD, USA) supplemented with EGM™-2 SingleQuots® supplement (Lonza, Walkersville, MD, USA). The cells were incubated at 37 °C and 5% CO2 saturation.
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Antioxidant Evaluation of Compounds

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ABTS radical cation (2,2′-azino-bis(3-ethylbenzothiazoline)-6-sulphonic acid), anthrone, DPPH (2,2-Diphenyl-1-picrylhydrazyl, (−)-epigallocatechin gallate (EGCG), gallic acid, sulforhodamine B (SRB), and trolox were obtained from Sigma Chemical (St. Louis, MO, USA). Folin–Ciocalteu reagent was purchased from Merck (Darmstadt, Germany). Dutasteride, finasteride, and minoxidil were from Wuhan W&Z Biotech (Wuhan, China). Agarose gel, Tris base, and 50X Tris/Acetic acid/EDTA (TAE) were from Bio-Rad Laboratories (Hercules, CA, USA). Follicle Dermal Papilla Cell Growth Medium Kit (cat no. C-26501) was from Promo Cell GmbH (Heidelberg, Germany). Antibiotic-antimycotic (100X; cat no.1 5240062), fetal bovine serum (FBS; cat no. 16000044), and Roswell Park Memorial Institute medium (RPMI-1640; cat no. 31800022) were from Gibco Life Technologies (Thermo Fisher Scientific, Waltham, MA, USA). Acetic acid, dimethyl sulphoxide (DMSO), trichloroAcetic acid, sulfuric acid, and other chemical substances were purchased from RCI Labscan (Bangkok, Thailand). All other chemicals were analytical-grade substances.
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4

Pangasius Skin Gelatin Hydrolysate Protocol

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LMWCP supplied by NEWTREE Co., Ltd., South Korea was prepared by spray drying the gelatin hydrolysate obtained by enzymatic degradation of gelatin derived from a skin of pangasius hypophthalmus using a protease and was standardized based on Gly-Pro-Hyp (3%) and tripeptide (≥15%) contents. hDPCs were purchased from PromoCell (Germany). The cells were maintained in an incubator with 5% CO2 at 37°C using a follicle dermal papilla cell growth medium kit (PromoCell) and subcultured upon reaching 70-80% confluency.
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5

Cultivation of Human Dermal Papilla and Keratinocytes

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The HFDPCs were purchased from PromoCell (Heidelberg, Germany) and cultured in Follicle Dermal Papilla Cell Growth Medium Kit (PromoCell) supplemented with 1% (v/v) penicillin/streptomycin (PS, Gibco BRL). Normal human epithelial keratinocytes (NHEKs) were purchased from PromoCell and cultured in keratinocyte growth medium 2 (PromoCell) supplemented with 1% (v/v) PS. The cells were incubated in 150-mm cell culture Petri dishes (Corning Inc., Corning, New York, NY, USA) until they reached 70–80% confluence. The cells were incubated at 37 °C and 5% CO2 saturation. The cell culture medium was changed every 3 days. Cells within 4 passages were used for the experiments.
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