Figure 6. PHH spheroids as a model for cholestatic and steatotic disease.

(A–C) Treatment with the cholestatic drug chlorpromazine (CPZ, 5 μM) caused significant accumulation (n = 3, p = 0.03) of the fluorescently-labelled bile acid derivative tauro-nor-THCA-25-DBD, which was associated with down-regulation of BSEP mRNA (C). (D,E) Cyclosporine A (CsA, 30 μM), a known inducer of steatosis in vivo⁴⁵ increased levels of neutral lipids (n = 3, p = 0.003). Strikingly, cells were fully protected by co-exposure with the anti-oxidant α-tocopherol (α-TOH, 10 μM) (n = 3, p = 0.009). Bile acid and lipid accumulation were quantified and normalized to spheroid size using CellProfiler software. All scale bars = 100 μm. *indicates p < 0.05, **indicates p < 0.01.