Fig. 2
From: 3D microfluidic liver cultures as a physiological preclinical tool for hepatitis B virus infection
3D PHH cultures are metabolically stable and exhibit functional tissue formation. a Comparison of cytochrome P450 gene (HNMT, CYP2A6, SLC22A1, CYP2A13, and GSTP1) mRNA expression between static 2D PHH cultures, 3D spheroid cultures, SACC PHH cultures, and 3D PHH cultures following 14 days of culture as well as freshly thawed PHH and HepG2 cells. b Longitudinal comparison of the mRNA expression profile of HNMT, CYP2A6, ABCB11, SLC22A17, CYP2A13, and GSTP1 as well as HBV receptors (NTCP, ASGPR) in HepG2 cells and freshly thawed PHH as well as static 2D and 3D PHH cultures. c Determination of CYP3A activity in six different hepatocyte donors 7 days following seeding in 3D PHH cultures as determined by CYP3A-Glo assay. d CYP450 activity in 3D PHH following 7, 14, or 21 days of culture time as determined by quantification of metabolites of Tacrine (CYP1A2), Diclofenac (CYP2C9), and Midazolam (CYP3A4) by quadrupole linear ion trap mass spectrometry. e Stable maintenance of the bile canaliculi marker DPPIV/CD26 in 3D PHH cultures as determined by Luminex. f Functionality of bile canaliculi as determined by CDFDA staining. g, h Ultrastructural analysis of 3D PHH by transmission electron microscopy after 20 days of culture. (Filled arrows: bile canaliculi, open arrows: hepatic microvilli, asterisk: tight junctions, L: lipid droplet). i Immunofluorescence microscopy of tight junction and hepatocyte markers (ZO-1, CD81, ITGB1, Connexin 32) in 3D PHH cultures following 26 days of culture. All data shown are mean ± SD of three to six independent experiments. Scale bars: white (200 μm), gray (2 nm), black (500 nm)
