Three-dimensional perfused human in vitro model of non-alcoholic fatty liver disease

Figure 2 Hepatocytes accumulate intracellular fat over time. Hepatocytes were cultured for 14 d under fat and lean conditions. Fat loading was measured by Oil Red O staining of microtissues, which were observed by microscopy (A), and quantified by absorbance at 510 nm and normalised to total cellular protein to give a relative fat content (B). Fat consumed by cells over 14 d of culture was calculated by analysing culture medium at each media change for the presence of free fatty acids using enzyme-based colorimetric assay (C). Hepatocytes were compared for albumin (D) and ALT/AST production (E). Each time point is a mean of 6 independent cultures ± SD (^aP < 0.01).