tib 202 nfkb luc2  (ATCC)

Journal: Biomedicines

Article Title: A Matrix-Free 3D Hepatocyte–Macrophage Co-Culture Spheroid Model for Dual Assessment of Lipid Accumulation and NF-κB-Mediated Inflammatory Activation Under Glucolipotoxic Stress

doi: 10.3390/biomedicines14040792

Figure Lengend Snippet: Morphology and metabolic activity of HepG2/THP-1 spheroids at different seeding densities. ( A ) Representative Live/Dead staining at Day 8 of co-culture spheroids seeded at 3000 (3k) or 6000 (6k) total cells/well. Viable cells are calcein-AM-positive (green) and dead cells are EthD-1-positive (red); corresponding nuclear counterstaining (DAPI, blue) is shown in the lower panels. Scale bars: 100 µm. ( B ) Metabolic activity during maturation (Days 3, 6, 8, and 10) measured by PrestoBlue™ fluorescence (Ex/Em 560/590 nm), normalized to the starting seeded cell number, and reported as normalized RFU. The graph shows one representative independent experiment; data are presented as mean ± SD of 8 technical replicate wells per condition. The experiment was repeated independently three times with comparable results. Two-way ANOVA (factors: time and seeding density) with Šídák’s multiple comparisons. Abbreviations: RFU, relative fluorescence units; EthD-1, ethidium homodimer-1; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: HepG2 and THP-1 NF-κB-Luc2 were authenticated by STR profiling (ATCC ® Lot number 70050519 and 70059144) and were mycoplasma-negative.

Techniques: Activity Assay, Staining, Co-Culture Assay, Fluorescence

Journal: Biomedicines

Article Title: A Matrix-Free 3D Hepatocyte–Macrophage Co-Culture Spheroid Model for Dual Assessment of Lipid Accumulation and NF-κB-Mediated Inflammatory Activation Under Glucolipotoxic Stress

doi: 10.3390/biomedicines14040792

Figure Lengend Snippet: Hepatocyte and macrophage marker expression during spheroid phenotypic remodeling. HepG2/THP-1 spheroids were harvested at Days 3, 6, 8, and 10 for bulk-spheroid RNA extraction followed by qPCR. ( A ) Hepatocyte-associated transcripts (ALB, AFP, CYP3A4) expressed as fold change versus 2D HepG2 monolayers (calibrator = 1.00). ( B ) Macrophage-associated transcripts (CD14, CD64, CD68, CD206, MARCO, TREM2) expressed as fold change versus 2D PMA-differentiated THP-1 cells (calibrator = 1.00). ( C ) Representative immunofluorescence at Day 8 showing albumin (ALB, green) and CD14 (red); nuclei counterstained with DAPI (blue). Scale bars: 100 µm. The graph shows one representative independent experiment; data are presented as mean ± SD of n = 4 biological replicates per condition, each biological replicate generated by pooling 2 spheroids. The experiment was repeated independently three times with comparable results. One-way ANOVA with Tukey’s multiple comparisons; groups not sharing letters differ significantly ( p < 0.05). Abbreviations: ALB, albumin; AFP, alpha-fetoprotein; CYP3A4, cytochrome P450 family 3 subfamily A member 4; CD, cluster of differentiation; MARCO, macrophage receptor with collagenous structure; TREM2, triggering receptor expressed on myeloid cells 2; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: HepG2 and THP-1 NF-κB-Luc2 were authenticated by STR profiling (ATCC ® Lot number 70050519 and 70059144) and were mycoplasma-negative.

Techniques: Marker, Expressing, RNA Extraction, Immunofluorescence, Generated