Paclitaxel taxol

Hair follicles were microdissected from human occipital scalp tissue obtained from the Crown Clinic, Manchester, donated by patients under informed consent. Hair follicles were cultured within 24‐well plates in 500 μl of hair follicle culture medium comprised of Williams E medium, 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM l‐glutamine, 10 μg/ml insulin and 10 ng/ml hydrocortisone (Philpott et al, 1994; Langan et al, 2015). Upon isolation, hair follicles were randomly allocated to a given treatment group and cultured ex vivo for the periods indicated in the presence of compound dissolved in DMSO or DMSO‐only (vehicle) at an equivalent concentration (max 0.01%). Compounds employed: 1 μM palbociclib (PD‐0332991) (Selleckchem, #S1116), 100 nM paclitaxel (Taxol) (Tocris, #1097) and 100 nM docetaxel (Taxotere) (Abcam, #ab141248). Following culture, hair follicles were embedded in cryoembedding matrix and frozen using liquid nitrogen and stored at minus 80°C.

Folic acid (FA), alginic acid sodium salt (ALG; viscosity 15.00–20.00 cps, 1% H₂O), trypsin from bovine pancreas (13,000 units/mg solid), gentamicin (50 µg/mL), dimethyl sulfoxide (DMSO), methylthiazoletetrazolium (MTT), 6-coumarin, ethylenediamine (ED; 98%), N-hydroxysuccinimide (NHS), N,N’-dicyclohexylcarbodiimide (DCC) were purchased from Sigma-Aldrich (Barcelona, Spain). Sodium hydroxide (NaOH), hydrochloric acid (HCl; 35%), trichloroacetic acid, ethanol absolute, ethylenediaminetetraacetic acid (EDTA), Tween^®80, triethylamine, anhydrous di-sodium hydrogen phosphate (Na₂HPO4), di-hydrogen potassium phosphate (KH₂PO4), diethylether, dichloromethane (DCM) and acetonitrile (HPLC-analysis grade) were supplied by Panreac (Madrid, Spain). Bovine serum albumin (BSA, Fraction V), sodium chloride (NaCl) and disodium hydrogen phosphate dehydrated were purchased from Merck (Barcelona, Spain). Fetal bovine serum (FBS), penicillin (50 U/mL), streptomycin (50 µg/mL) and 0.05% trypsin/0.53 mM EDTA were purchased from Invitrogen Life Technologies, Grand Island, NY. Dubecco’s Modified Eagle Medium was purchased from Lonza Bioscience (Bornem, Belgium). Paclitaxel (Taxol) was supplied by Tocris Bioscience (Mw 889.95 > 99%). All water used in the assays was Millipore Milli Q grade.