Extended Data Fig. 7: Validating the maturation of in vitro derived hepatocytes using an inducible expression culture system.
From: Single-cell atlas of human liver development reveals pathways directing hepatic cell fates
a, Schematic of the experimental design for doxycycline (DOX) induction of key transcription factors during hepatocyte-like cell differentiation from hPSCs. HiPSC cells were edited to include the transcription factor in an inducible cassette, which was upregulated at day 15 of differentiation using DOX and continued to be expressed until day 23 when the HLCs were assays for maturation via qPCR; created with BioRender.com. b, QPCR analysis showing expression of NFIX, NFIA or CAR by day 23 hepatocyte-like cells (HLCs) that were induced to express their respective transcription factors since day 15 of differentiation using a doxycycline-inducible expression system (n≥3). The induced expression levels are compared to primary foetal and adult human liver, showing that expression is within physiological bounds. c, In addition to the increase in mature albumin expression and the decrease of foetal AFP expression, qPCR shows the increase in a wide array of functional and metabolic hepatocyte markers, thus, indicating maturation due to temporally-relevant expression of key transcription factors (n = 6). Data are presented as mean values + /- SEM; Mann-Whitney unpaired two-tailed t-tests.
