Gibco horse serum hs

The pheochromocytoma cell line (PC 12), derived from rat adrenal medulla, was purchased from the American Type Culture Collection (ATCC, Manassas, Virginia, United States of America). PC 12 cells were cultured in complete Gibco™ RPMI medium (Thermo Fisher Scientific, Scoresby, Victoria, Australia) supplemented with 10% Gibco™ horse serum (HS; Thermo Fisher Scientific), 5% Gibco™ foetal bovine serum (FBS; Thermo Fisher Scientific) and 1% Gibco™ penicillin/streptomycin (PS; Thermo Fisher Scientific). Supplements were stored as aliquots at −20 °C. Stock solutions of PC 12 cells were prepared in a medium containing 90% FBS and 10 % DMSO and stored in liquid nitrogen. The cells were maintained at a temperature of 37 °C under a 5% CO₂ atmosphere in a 95% humidified incubator. The medium was changed every two days and passaged accordingly when the confluence reached 90%.

The pheochromocytoma cells (PC12) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA)) and were cultured in complete Gibco™ RPMI medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% Gibco™ horse serum (HS, Thermo Fisher Scientific, Waltham, MA, USA), 5% Gibco™ foetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA), and 1% Gibco™ penicillin/streptomycin (PS, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C and 5% CO₂ in a 95% humidified incubator. The medium was changed every two days and passaged accordingly when cell confluence reached 90%.
For each independent experiment, PC12 cells were seeded at a density of 10,000 cells per 100 µL on AR-Ti and HTE-Ti samples. After 1-, 5-, and 7-day incubation periods, the samples were prepared for imaging as described in the following sections. Cell proliferation and total protein count assays were performed after 1 day of incubation to study the attachment patterns. All experiments were approved under the Swinburne Biosafety Project 2014/SBC01.