Figure 3.
Hepatocellular Differentiation of Canine Hepatic Organoids
(A) Dot plots show fold change of gene expression of organoids cultured in DM from different passage numbers (P3, P12, and P22) toward organoids cultured in EM in the same passage (n = 3 biological replicates). Expression levels in primary hepatocytes are shown in Figure S3.
(B) Hepatic enzymes and albumin secretion of organoids in DM were measured and presented as fold change toward organoids in EM (n = 3 biological replicates). Albumin secretion in organoids was also compared to HepG2, Huh7, and primary hepatocytes (Hep.). Data presented as average ± SD.
(C) Cyp3a activity was measured in organoids cultured in DM and presented as fold change toward organoids in EM (n = 3 biological replicates). The graph compared Cyp3a activity of organoids in DM to HepG2, Huh7, and Hep. Data presented as average ± SD.
(D) Representative images of immunohistochemical stainings of organoids cultured in EM and DM. Organoids in DM lose cholangiocyte markers such as SOX9 and gain mature hepatocyte markers such as ZO1 and MRP2.
(A)–(D) show the results from one organoid isolation. The experiment has been performed on organoids derived from at least three more different isolations and in different passages, which all show similar results.