ASSESSMENT OF LIVER-FUNCTION IN PRIMARY CULTURES OF HEPATOCYTES USINGDIETHOXY(5,6)CHLOROMETHYLFLUORESCEIN AND CONFOCAL LASER-SCANNING MICROSCOPY

A method is presented which can be used to assess the function of hepa tocytes in complex culture configurations without disrupting the integ rity of the cell environment. It utilises a fluorescent probe for cyto chrome P450 dependent mixed function oxidase (MFO) activity, diethoxy (5,6) chloromethylfluorescein, and confocal laser scanning microscopy. The MFO activity of individual cells in primary cultures of intact he patocytes can be detected in situ, and quantified by image analysis. T his may be a valuable means of monitoring the effect of culture condit ions on the function of bioartificial liver devices, and could be used to assess the need for effective oxygenation of cells, the influence of shear stress and of exposure to patient serum during clinical use.