FIGURE 3.

Differentiation of bipotent HepaRG cells to hepatocytes and cholangiocytes and differentiation‐induced expression of lnc‐RHL. A, Undifferentiated HepaRG cells are expanded in growth medium for 14 days yielding bipotent cells, then grown in differentiation medium for an additional 14 days, yielding mixed cultures of both hepatocytes and cholangiocytes. B, RT‐PCR assays from independent cultures (triplicate) at day 14 (undifferentiated) and day 33 (differentiated). Note robust induction of lnc‐RHL expression upon differentiation. C, Morphology of HepaRG cultures at day 14 (undifferentiated) and day 33 (differentiated). Note that day 33 cultures consist of a monolayer of cholangiocytes upon which 3‐dimensional colonies of hepatocytes occur