Journal: bioRxiv

Article Title: Novel hepatocyte-like liver organoids recapitulate crucial mature hepatic functions

doi: 10.1101/2024.10.29.620824

Figure Lengend Snippet: (A) Schematic overview of the HeLLO establishment protocol. (B) HeLLO brightfield imaging showed similar morphology in the expansion medium (EM) compared to ICOs, while collapsing into distinct dense organoids in the differentiation medium (DM). (C) Immunofluorescence staining showed strong expression of liver proteins albumin, CYP3A4, E-cadherin and CYP2E1 in HeLLO-DM but not in ICO-DM (both at differentiation day 8). (D) CellNet liver classification of transcriptomic data resulted in a higher liver classification in HeLLO-DM compared to ICO-DM. (E) Heatmap analysis of transcriptomic data indicated that HeLLOs gain hepatocyte phenotypes while losing some cholangiocyte markers upon differentiation. Additional liver samples were integrated using HLCompR.

Article Snippet: Cryopreserved primary human hepatocytes (PHHs; BeCytes Biotechnologies) were thawed and resuspended in hepatocyte plating medium: William’s E medium supplemented with 2 mM GlutaMAX, 1% Non-Essentials Amino Acid Solution (Gibco, 11140050), 100 U/ml PenStrep, 1x ITS+ Premix (Corning, 354352), 1 mM N-acetylcysteine, 100 nM Dexamethasone, and 5% FBS (Gibco, 26400044).

Techniques: Imaging, Immunofluorescence, Staining, Expressing

Journal: bioRxiv

Article Title: Novel hepatocyte-like liver organoids recapitulate crucial mature hepatic functions

doi: 10.1101/2024.10.29.620824

Figure Lengend Snippet: (A) UMAP visualization of one donor-matched ICO and HeLLO samples in expansion and differentiation media. (B) UMAP showing sub clusters of ICO-EM (IE), ICO-DM (ID), HeLLO-EM (HE), and HeLLO-DM (HD) samples. (C) UMAP showing SingleR classification scores for adult hepatocytes from the human liver development atlas . (D) UMAP showing expression of hepatocyte (top) and cholangiocyte (bottom) marker genes. Color indicates normalized expression data. (E) Violin plots of the HeLLO-DM cluster HD3 and hepatocytes from the human liver cell atlas showing the normalized expression of hepatocyte (top) and cholangiocyte (bottom) marker genes. (F) Trajectory analysis on the HeLLO-EM and HeLLO-DM samples. White line indicates the inferred trajectory, the color bar indicates the pseudotime, and color shading in the background represents an approximation of the clusters. (G) Dot plots showing the expression level (color) and percentage of cells (dot size) expressing the proliferation, cholangiocyte, secreting cholangiocyte, and hepatocyte markers on each cluster. This shows clear differences between the clusters with HD3 being the most hepatocyte-like. (H) UMAP showing the module score for upregulated and downregulated gene sets in transitional liver progenitor cells (TLPCs) against biliary epithelial cells (BECs) from Pu et al. (2023).

Article Snippet: Cryopreserved primary human hepatocytes (PHHs; BeCytes Biotechnologies) were thawed and resuspended in hepatocyte plating medium: William’s E medium supplemented with 2 mM GlutaMAX, 1% Non-Essentials Amino Acid Solution (Gibco, 11140050), 100 U/ml PenStrep, 1x ITS+ Premix (Corning, 354352), 1 mM N-acetylcysteine, 100 nM Dexamethasone, and 5% FBS (Gibco, 26400044).

Techniques: Expressing, Marker