Measuring mitochondrial function and glycolytic flux in 3D cell cultures

Conn Carey, James Hynes Luxcel Biosciences 09/2014

Metabolic processes like glycolysis and respiration are essential functions that shed light on cellular health and should be monitored in response to potential drugs. The MitoXpress® Xxtra HS assay from Agilent Technologies uses an O2 sensitive fluorophore for measuring oxygen consumption which correlates with mitochondrial function. The pH-Xtra™ assay uses a pH-sensitive fluorophore which detects acidification due to glycolysis-related release of lactate. Recently, 3D cell cultures aim at bridging between conventional cell culture at in vivo studies.

This application note shows the monitoring of metabolism in RAFT™ 3D cultures of A549 lung cancer cells with the FLUOstar® Omega or CLARIOstar® microplate reader.

Increases in oxygen consumption were determined for increasing cell densities as well as upon addition of the mitochondrial uncoupler FCCP. Furthermore, modulation of lactate production by addition of oxamate or antimycin led to lower or higher acidification, respectively, as detected with pH-Xtra on FLUOstar® Omega.

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