Fig. 1. Experimental procedure to elucidate the manner of hepatic stem cell differentiation. The Met⁺ CD49f^+/low Kit^– CD45^– TER119^– cells isolated from E13.5 fetal mouse livers were cultured clonally in 96-well plates. Cells that expanded in culture and produced hepatocytes and cholangiocytes as descendants while maintaining primitive stem cells undergoing self-renewing divisions were used to analyze differentiation status. We chose three independent stem cell clones for examination. Following transfection of the ALB enhancer/promoter-EGFP construct, cells were examined by using FACS and quantitative PCR after they had been cultured with several GFs or ECMs. Scale bar: 100 µm.