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. Author manuscript; available in PMC: 2021 Oct 14.
Published in final edited form as: Adv Funct Mater. 2020 Jun 8;30(48):2000893. doi: 10.1002/adfm.202000893

Figure 4. Organoids differentiated into functional hepatocyte-like cells in PIC.

Figure 4

Organoids were differentiated in Matrigel, 1k PIC-LEC, or 5k PIC-LEC for 8 days (N = 4). a) Light microscopy images of organoids showing that differentiated organoids became denser and darker compared to EM organoids in all hydrogels. b) Gene expression of stem cell and hepatocyte markers in differentiated organoids from four independent donors. Transcriptional levels of leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5), albumin (ALB), cytochrome p450 3A4 (CYP3A4), and multidrug resistance-associated protein 2 (MRP2) were determined by qRT-PCR and compared to cryopreserved human hepatocytes. Hepatocyte functionality of differentiated organoids in the different hydrogels was assessed. Four different donors were analyzed in independent experiments. d) Rh123 transport was determined as read-out for MDR1 activity. Verapamil was added as an inhibitor of MDR1 function. c) Albumin concentrations and e) GLDH levels in cell lysates were measured after incubation in DM for 24 h. Comparison was normalized to total protein concentrations. Graphs indicate mean ± SD. f) Ammonium elimination from the culture medium was determined as read-out for hepatocyte functionality. *indicates p-value ≤ 0.05.