Journal: Scientific Reports

Article Title: Expression and function of CYP enzymes and hepatobiliary transporters in an improved long-term sandwich culture hepatocyte model

doi: 10.1038/s41598-025-33332-9

Figure Lengend Snippet: Activities of CYP enzymes in SPHH, SPHH-5C and PHH-5C hepatocyte cultures up to 21 days. The following CYP enzymes are presented: ( a ) CYP1A2, ( b ) CYP2B6, ( c ) CYP2C8, ( d ) CYP2C9, ( e ) CYP2C19, ( f ) CYP2D6 and ( g ) CYP3A4. D0 values are determined in the cell suspension before seeding. Representative graphs from single-donor hepatocytes (lot: IRZ). Data is presented as Mean ± SD ( n = 3 of each condition). Two-way ANOVA with Tukey’s multiple comparison tests ( p < 0.05).

Article Snippet: The applied TaqMan probes (ThermoFisher) were the following: 18 S (Hs99999901_s1), CYP1A2 (Hs00167927_m1), CYP2B6 (Hs04183483_g1), CYP2D6 (Hs04931916_gH), CYP2C8 (Hs00426387_m1), CYP2C9 (Hs04260376_m1), CYP2C19 (Hs00426380_m1), CYP3A4 (Hs00604506_m1), OTP1B1 (Hs00272374_m1), NTCP (Hs00914890_m1), OCT1 (Hs00427552_m1), OAT2 (Hs00198527_m1), BCRP (Hs01053790_m1), BSEP (Hs00994805_m1), MRP2 (Hs00960489_m1), MDR1 (Hs01070654_g1), MDR3 (Hs00240956_m1).

Techniques: Suspension, Comparison

Journal: Bioactive Materials

Article Title: Three-dimensional bioprinted hiHeps hepatorganoids with enhanced hepatic functions for the treatment of liver failure and promotion of liver regeneration

doi: 10.1016/j.bioactmat.2025.12.024

Figure Lengend Snippet: Zonation characterization of 3D bioprinted human hepatocyte organoids. A. Representative live/dead assay of hiHep cells in 2D and 3D culture. Scale bar: 200 μm. B. Quantification of cell viability of hiHep cells in 2D and 3D culture (n = 3 per group). C. Cell viability of 2D cultured hiHeps and 3DP-HHO assessed by CCK-8 assay (n = 3 per group). D. Morphological comparison of 2D cultured hiHeps, 3D bioprinted hiHeps organoid, and 3DP-HHO after 1 week/month of culture (Red arrow: liver cord-like structures formed via self-assembly, white arrow: hepatocyte spheroids spontaneously formed by hiHeps). Scale bars: 50 μm. E. Volcano plot of differentially expressed genes in 3DP-HHO compared to 2D cultured hiHeps; key hepatic genes are highlighted. F. KEGG enrichment analysis of up-regulated genes showing biological processes associated with hepatic function. G. Heatmap of zonation marker gene expression (pericentral vs. periportal hepatocyte landmarks) across HepRG, hiHeps, 3DP-HHO, and primary human hepatocytes (PHH). H. Schematic diagram of zonation map illustrating the spatial gene expression of pericentral (red) and periportal (green) markers in healthy liver. I. Representative immunofluorescence images and quantification of CYP3A4 (red), HAL (green), and DAPI (blue) in healthy mouse liver, 2D-cultured hiHeps and 3DP-HHO, demonstrating spatially organized metabolic domains. (White arrow: CYP3A4-positive zones, indicating pericentral hepatocytes, white pentagram: HAL-positive zones, indicating periportal hepatocytes). Scale bars: mouse liver, 100 μm; 3DP-HHO, 25 μm.

Article Snippet: CYP3A4 Mouse mAb, 1:500, 67110-1-Ig, Proteintech, China.

Techniques: Live Dead Assay, Cell Culture, CCK-8 Assay, Comparison, Marker, Gene Expression, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Ammonium Hydroxide Enhancement of Dietary Protein in High-Fat Diets Modulates Liver Metabolism Signaling in a Sex- and Age-Dependent Manner in C3H/HeJ Mice

doi: 10.3390/ijms27010403

Figure Lengend Snippet: Predicted AHE-induced changes to hepatic metabolite levels and proteins associated with ammonia metabolism. ( A ) A simplified representation of the Krebs cycle. Arrows indicate predicted levels of metabolites that may be affected by AHE. ( B ) The role of β-catenin in the typical high-affinity low-capacity clearance of ammonia in hepatocytes utilizing GS. Additionally, CYP3A4 is predicted to change, through a yet undetermined mechanism potentially associated with GS (dashed, double-headed arrow). Blue arrows = predicted decreases. Orange arrows = predicted increases.

Article Snippet: Primary antibodies and their incubation conditions were: β-catenin at RT for 1 h (1:10,000; ProteinTech, Rosemont, IL, USA: 66379-1-Ig), GS at 4 °C overnight (1:1000; NovusBio, Centennial, CO, USA: NB110-41404), CYP3A4 at 4 °C overnight (1:10,000; ProteinTech, Rosemont, IL, USA: 67110-1-Ig) and HSP90 (housekeeping) at RT for 1 h (1:1000; Cell Signaling Technology, Danvers, MA, USA: C45G5).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Ammonium Hydroxide Enhancement of Dietary Protein in High-Fat Diets Modulates Liver Metabolism Signaling in a Sex- and Age-Dependent Manner in C3H/HeJ Mice

doi: 10.3390/ijms27010403

Figure Lengend Snippet: Densitometric measures of relative expression of key proteins linked with ammonia metabolism. ( A ) Female relative expression as mean ± SEM at 6, 12 and 18 months, respectively. ( B ) Male relative expression as mean ± SEM at 6, 12, and 18 months, respectively. ( C ) Representative Western blots of each protein at 18 months in females. ( D ) Representative Western blots of each protein at 18 months in males. Proteins analyzed: top: β-catenin (mw ~90 kDa, ProteinTech, 1:10,000), second: GS (mw ~42 kDa, NovusBio, 1:1000), and third: CYP3A4 (mw ~51–55 kDa, ProteinTech, 1:10,000). Bottom: HSP90 (mw ~90 kDa) used as the reference. ( E , F ) CYP3A4 Western blots from each timepoint for females and males, respectively. Significance between diet groups is indicated at p -value < 0.05 by different letters (i.e., result A is significantly different from result B). Samples were not available for assessment in HFBN females at 6 months; the third replicate in HFB females at 12 months was not available due to an issue with protein lysate stability; the third replicate for HFC males at 18 months was not available due to lack of non-cancerous liver tissue for lysis.

Article Snippet: Primary antibodies and their incubation conditions were: β-catenin at RT for 1 h (1:10,000; ProteinTech, Rosemont, IL, USA: 66379-1-Ig), GS at 4 °C overnight (1:1000; NovusBio, Centennial, CO, USA: NB110-41404), CYP3A4 at 4 °C overnight (1:10,000; ProteinTech, Rosemont, IL, USA: 67110-1-Ig) and HSP90 (housekeeping) at RT for 1 h (1:1000; Cell Signaling Technology, Danvers, MA, USA: C45G5).

Techniques: Expressing, Western Blot, Lysis