Thiazolyl blue formazan mtt

1-(4, 5-Dimethylthiazol-2-yl)-3, 5-diphenylformazan or thiazolyl blue formazan (MTT), finasteride and hydrocortisone were purchased from Sigma-Aldrich (St. Louis, USA). Fetal bovine, 2.5% trypsin serum, penicillin/streptomycin, phosphate buffer solution pH 7.4 were obtained from Gibco (Auckland, New Zealand). Dimethyl sulfoxide or DMSO (Analytical grade) was purchased from Labscan (V.S. Chem House, Thailand). Mouse IL-1β ELISA Instant Kit was purchased from e-bio sciences (Bender-med systems Gmblt, Vienna, Austria). Follicle dermal papilla cell growth medium was purchased from PromoCell (Heidelberg, Germany).

Baculovirus titres in the VLP vaccine doses were determined by plaque assay on Sf9 cells with minor modifications as described in [24] . Briefly, the assay was carried out in 6-well plates in duplicates. After seeding 1 × 10⁶ cells per well, the cells were allowed to attach to the surface, medium was removed and 200 μL of the diluted VLP vaccine formulations (10-fold dilutions in TNM-FH unsupplemented) were added and incubated for 1 h at 27 °C with periodic shaking. After infection, the samples were removed and cells were overlaid with 2 mL of a solution containing 1% agarose in TNM-FH, 10% (v/v) foetal bovine serum, Penicillin–Streptomycin antibiotic mixture pre-warmed to 37 °C. The plates were incubated at 27 °C for 6 days and plaques were counted after live-cell staining with 200 μL of 5 mg/mL Thiazolyl blue formazan MTT (Sigma, St. Louis, MO) for 3–4 h.