concentrations
ranging up
until (3)
360 mM in moderate elevated though physiological concentrations,
and (4) 720 mM in high pathologic and
potentially cytotoxic metabolic conditions.
Stearic acid (SA), palmitic acid (PA), and oleic acid (OA;
Sigma–Aldrich) were dissolved in a stock solution of pure
ethanol at concentrations of 25, 150, and 200 mM,
respectively. These ethanol stock solutions were vortex
mixed for 4 minutes and diluted to obtain the desired final
concentration in the culture medium. Before use, all media
were vigorously shaken and filter sterilized under aseptic
conditions. Non-esterified fatty acid solvability was
checked in all NEFA medium samples before exposure to
determine whether the intended concentrations wereactually achieved. Final NEFA concentrations were
measured at the “Algemeen Medisch Labo” (Antwerp,
Belgium), using commercial photometric assays, RX Daytona
(Randox Laboratories). Measurements were carried
out according to the manufacturers’ instructions