Research Article

Fine Tuning of Hepatocyte Differentiation from Human Embryonic Stem Cells: Growth Factor vs. Small Molecule-Based Approaches

Figure 6

Functional analysis of hepatocytes differentiated from various DE derivation methods. (a) Undifferentiated H9 cells, HepG2 cells, and H9-derived DE cells using growth factors (growth factors/NaB/DMSO (5 d)) and (growth factors/NaB (5 d)) and CHIR (3 μM CHIR (6 d)) protocols and differentiated into hepatocytes with either Carpentier’s (D18, growth factors/NaB/DMSO (5 d)/Carpentier’s and D18, growth factors/NaB (5 d)/Carpentier’s) or Touboul’s (D18, growth factors/NaB/DMSO (5 d)/Touboul’s and D18, growth factors/NaB (5 d)/Touboul’s) or CHIR (3 μM CHIR (6 d)/Touboul’s) maturation media were assayed for lipid accumulation and glycogen storage using Oil Red O staining and periodic acid-Schiff (PAS) staining, respectively. (b) CYP3A4 activity was measured within intact cells (same as described in (a)) using a nonlytic cell-based assay by employing Luciferin-IPA (isopropyl acetal) as the substrate. The bars represent relative luminescence units (RLU), and the significance in the luminescence ( values) for all cell types is derived relative to the values for undifferentiated H9 cells. and .