Pitavastatin Cytotoxicity on PD-GSCs

PD-GSCs were harvested from a T75 flask and passaged into replicate T75 flasks for either pitavastatin (6μM) or vehicle (DMSO) treatment (2.0e6 cells/flask). Concomitantly, a portion of those PD-GSCs were used to inoculate laminin-treated 96 well plates for drug-dosing analysis (see IC₅₀Analysis section). On D4, PD-GSCs were harvested using Accutase (1mL/25cm²) as described previously. Cell suspensions were spun at 1000rpm (193g) for five minutes. Cell pellets were then resuspended with serum-free culture media (200,000 cells/mL) to inoculate 96 well plates (100μL/well, 20,000 cells/well) for subsequent IC₅₀ determination. PD-GSCs were incubated in serum-free culture media in 96 well plates for 48 hours to allow for cell attachment prior to replacing spent media with serum-free media with pitavastatin (or vehicle). Treated cells were incubated at 37°C for four days. Following the four-day treatment, cell viability was measured via MTT assay as described below.