In vitro differentiation of hepatoblasts into hepatocytes

We differentiated primary mouse E14.5 Dlk1+ hepatoblasts in vitro into hepatocyte-like cells that recapitulated morphological features of hepatocytes. To assess the level of differentiation, we performed RNA-seq analysis and compared gene expression profiles of hepatobalsts, in vitro differentiated hepatocytes and mature hepatocytes isolated from adult mouse livers as a control. Hepatoblasts, in vitro differetniated hepatocytes and mature hepatocytes were collected in 4 biological replicates. Hepatoblasts were isolated based on the Dlk1 surface marker using MACS (Tanimizu et al., 2003, JCS). The isolated cells were cultured on fibronectin coated plates in optimized Differentiation media for 4 days (Belicova et al., 2021, JCB). The adult mouse liver hepatocytes were isolated according to a published colagenase perfusion protocol (Klingmüller et al., 2006, IEE Proceedings: Systems Biology).