Skip to main content
. Author manuscript; available in PMC: 2020 Jan 28.
Published in final edited form as: Differentiation. 2019 Jan 28;105:54–70. doi: 10.1016/j.diff.2019.01.005

Fig. 1. hESC cultured with FHB-CM shifted to epithelial morphology resembling hepatocytes.

Fig. 1.

(A) Phase contrast microscopy: Undifferentiated hESC under lower and higher magnifications as indicated (top); hESC treated with growth factors and other additives alone (GFs) after 3d and 14d (second from top); eFHLC derived from hESC cultured with hTERT-FH-CM for 3d or 14d (second from bottom); and eFHLC after first passage (P1) compared with primary fetal hepatocytes (FH) in initial culture (P0) (bottom panel). In hESC cultured with GFs, cells did not develop epithelial morphology (inset, higher magnification shows cells with morphology of hESC). By contrast, culture of hESC with hTERT-FH-CM led to morphological alterations within 3d. By 13-14 d, these eFHLC universally exhibited epithelial morphology – inset, higher magnification including cells with binucleation – a feature of hepatocytes. Original magnification ×100-400; scale bars, 15 μm. (B) Transmission EM of eFHLC and freshly-isolated fetal hepatocytes without culture (FH-PP). In these cell types, nucleus was often to the side with vesicles or microperoxisomes characteristic of hepatocytes (scale bar, 1 μm).