human normal hepatocytes  (ATCC)

Journal: Journal of Cellular and Molecular Medicine

Article Title: Regulation of Autophagy and Metabolism in Hepatocellular Carcinoma: Involvement of Wnt‐β‐Catenin Pathway

doi: 10.1111/jcmm.71070

Figure Lengend Snippet: Inhibition of cell viability and colony formation, and induction of apoptosis by riluzole. (A) Expression of stem cell markers in HCC CSCs. CSCs were stained with CD44 and CD133 antibody and the expression of CD44 and CD133 was measured by immunocytochemistry. (B, C) HCC cell lines (HepG2 and Hep3B), cancer stem cells (CSCs), and human normal hepatocytes were seeded in 96‐well plates, treated with or without riluzole (0–40 μM) for 72 h, and cell viability was measured by CellTiter‐Glo Luminescent Cell Viability Assay (Promega). (D) Colony formation. HCC cell lines (HepG2, Hep3B, SNU‐382 and SNU‐475) and CSCs were treated with riluzole (0–20 μM). Number of colonies formed at 21 days was counted under a microscope. (E) Normal human hepatocytes, HCC cell lines (HepG2 and Hep3B) and CSCs were treated with riluzole (0–20 μM) for 72 h. Apoptosis was measured by TUNEL assay. Data represent mean ± SD ( n = 4). *, # and % = significantly different from control and each other; p < 0.05.

Article Snippet: Human normal hepatocytes were purchased from ATCC.

Techniques: Inhibition, Expressing, Staining, Immunocytochemistry, Cell Viability Assay, Microscopy, TUNEL Assay, Control

Journal: Frontiers in Pharmacology

Article Title: Ultrafine garlic powder alleviates non-alcoholic steatohepatitis by inhibiting hepatocyte ferroptosis and modulating ERK-dependent oxidative stress

doi: 10.3389/fphar.2025.1711917

Figure Lengend Snippet: UGP ameliorates palmitatic acid-induced lipotoxic hepatocyte injury. Effects of palmitatic acid stimulation and Fer-1, UGP and TGP treatment of on human normal hepatocyte THLE2 cell viability was examined using CCK8 assay (A) and LDH assay (B) (n = 4). (C) Effects of palmitatic acid stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell damage was determined by ALT assay (n = 4). (D) Cell death assay was performed using Annexin V/PI staining (n = 4). Annexin V + PI − , Annexin V + PI − , and Annexin V + PI + cells were considered dead cells. Representative fluorescent images and quantification of DCFDA (2′,7′-dichlorofluorescein diacetate) (E) and Mito-Tracker Red CMXRos (F) staining for ROS and mitochondrial membrane potential detection (scale bar = 100 μm, n = 4). (G) Flow cytometry analysis of lipid peroxidation by Liperfluo staining on THLE2 cells. (H) Representative immunofluorescent images of ACTA2 +ve and COL1A1 +ve cells (scale bar = 100 μm). Data are shown as box-and whisker with median (middle line), 25th–75th percentiles (box), and min-max values (whiskers), one-way ANOVA with Tukey’s correction. (ALT: Alanine transaminase; BSA:Bovine serum albumin; Fer-1: Ferrostatin-1; PAL: Palmitic acid; ROS: Reactive oxygen species; TGP: Traditional garlic powder; UGP:Ultrafine garlic powder).

Article Snippet: Human normal hepatocytes (THLE2) and hepatic stellate cells (HSCs) (LX-2) were purchased from the American Type Culture Collection (ATCC) and cultured in BEGM kit medium and RPMI medium supplemented with 10% FBS, respectively.

Techniques: CCK-8 Assay, Lactate Dehydrogenase Assay, Staining, Membrane, Flow Cytometry, Whisker Assay

Journal: Frontiers in Pharmacology

Article Title: Ultrafine garlic powder alleviates non-alcoholic steatohepatitis by inhibiting hepatocyte ferroptosis and modulating ERK-dependent oxidative stress

doi: 10.3389/fphar.2025.1711917

Figure Lengend Snippet: UGP significantly ameliorates erastin-induced hepatocyte ferroptosis. Effects of erastin stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell viability was examined using CCK8 assay (A) and LDH assay (B) (n = 4). (C) Effects of erastin stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell damage was determined by ALT assay (n = 4). (D) Cell death assay was performed using Annexin V/PI staining (n = 4). Annexin V + PI − , Annexin V + PI − , and Annexin V + PI + cells were considered dead cells. (E) Flow cytometry analysis of cellular Fe 2+ levels by FerroOrange staining on THLE2 cells. Representative fluorescent images and quantification of DCFDA (2′,7′-dichlorofluorescein diacetate) (F) and Mito-Tracker Red CMXRos (G) staining for ROS and mitochondrial membrane potential detection (scale bar = 100 μm, n = 4). (H) Flow cytometry analysis of lipid peroxidation by Liperfluo staining on THLE2 cells. Data are shown as box-and whisker with median (middle line), 25th-75th percentiles (box), and min-max values (whiskers), one-way ANOVA with Tukey’s correction. (ALT: Alanine transaminase; ERA: Erastin; Fer-1: Ferrostatin-1; ROS: Reactive oxygen species; TGP: Traditional garlic powder; UGP: Ultrafine garlic powder).

Article Snippet: Human normal hepatocytes (THLE2) and hepatic stellate cells (HSCs) (LX-2) were purchased from the American Type Culture Collection (ATCC) and cultured in BEGM kit medium and RPMI medium supplemented with 10% FBS, respectively.

Techniques: CCK-8 Assay, Lactate Dehydrogenase Assay, Staining, Flow Cytometry, Membrane, Whisker Assay