A cell concentration of 2 105 cell/cm3 were seeded in petri
dishes (3 cm3) and allowed to adhere for 24 h at 37 C using a
humidity of 5% CO2. The following day, the old media was discarded
and the cells were treated with complexes 1 and 2, using
a final concentration of 10 lM. This concentration was achieved
by the preparation of 1 mM stock solution of the silver(I) complexes
in culture grade DMSO. The complexes’ stock solution
(10 lL from the 1 mM) was added to the media, to give a final concentration
of 10 lM in 1 mL of treatment media, with 1% DMSO.
The cells were incubated for 24 h at 37 C in a 5% CO2 environment.
The control treatments include; untreated control, vehicle control
(cells treated with 1% DMSO), apoptotic control (100 lM cisplatin)
and necrotic control (5% H2O2). The % viability of the cells after
treatment was determined by the alamarBlue viability assay. This
assay involves a 100 lL suspension of treated cells in triplicate in a
96-well plate. The alamarBlue dye (10 lL) was added in the
absence of light, followed by an incubation for 2–3 h at 37 C and
5% CO2 humidity. The fluorescence of the samples were measured
using a Synergy HT Multi-Detection Microplate reader at an excitation
and emission wavelengths of k 530 nm and k 590 nm, respectively.
The % viability was determined based on the fluorescence of
the untreated control.