2.4. PC12 and IMR32 cell culture and treatment
Rat PC12 pheochromocytoma cells and human IMR32 neuroblastoma
cells were purchased from Bioresource Collection and
Research Center (BCRC, Hsinchu, Taiwan). PC12 cells were
cultured in Roswell Park Memorial Institute (RPMI) 1640 medium
with 10% horse serum (HS), 5% fetal bovine serum (FBS), and
2 mM L-glutamine at 37 C in a 5% CO2 humidified environment.
IMR32 cells were cultured in minimum essential media (MEM)
medium with 10% FBS at 37 C in a 5% CO2 humidified environment.
Poly lysine was prepared by adding 50 mL sterile tissue
culture grade water to 5 mg poly lysine. Coated cell culture surface
with 1 mL/25 cm2 culture surface. After 15 min removed
solution by aspiration and thoroughly rinsed surface with sterile
tissue culture grade water. Finally, the cell culture was sterilized
under the hood and UV light for 10e15 min. PC12 cells were
seeded on poly-L-lysine-coated plates and cultured for 24 h, followed
by culturing in the medium containing 100 ng/mL nerve
growth factor for 6 days. Differentiated PC12 cells were treated
with or without aggregated Ab1e40 peptide in the presence of CAE
(0, 25, 50, or 100