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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 5. Long-term expansion of organoids in PIC.

Figure 5

Organoids from two different donors were cultured for 12 weeks in Matrigel, 1k PIC-LEC, and 5k PIC-LEC with weekly passaging. a) Light microscopy pictures show that organoids maintained their morphological phenotype during all passages in Matrigel, 1k PIC-LEC, and 5k PIC-LEC. b) qRT-PCR analysis of the stem cell marker LGR5 shows stable expression levels at passage 2, 6, 10, and 14 in all different hydrogels. c) mRNA sequencing on organoids from two independent donors in EM and after differentiation (day 8 in DM). The 100 most significant differentially-expressed genes in DM versus EM are displayed in a heatmap. Note that EM samples from both donors and all three hydrogels show a very similar expression pattern and that all DM samples from both donors and all three different hydrogels are similar. A full list of genes is provided as Table S1, Supporting Information. d) Gene expression of hepatocyte markers in differentiated organoids from two independent donors during long-term culture in Matrigel or PIC hydrogels. Transcriptional levels of hepatocyte markers ALB, CYP3A4, and MRP2 were determined by qRT-PCR and compared to their respective EM controls (N = 2).