Figure 7: Modulation of Notch signalling influences the differentiation of bipotent HepaRG cells.

HepaRG cells subjected to differentiation conditions formed (Ai) tight colonies that immunostained positive for the hepatocyte markers (Aii) HNF4α and (Aiii) Albumin whereas the surrounding cells immunostained positive for the biliary epithelial marker (Aiv) CK19 and (Av) β-Catenin. The interface between colonies and surrounding cells immunostained positively for (Avi) NICD. Bipotent HepaRG cells were transduced with lentiviral constructs expressing Numb or Jag1/GFP then subjected to differentiation conditions. We observed a greater area of hepatocyte-like colonies in (Bi) HepaRG-Numb cells compared to (Bii) HepaRG-Jag1/GFP cells as quantified by total area ImageJ software. In HepaRG-Jag1/GFP cultures dual immunocytochemistry revealed that (Ci) GFP and CK7 co-localized but (Cii) GFP and Albumin never co-localized.