Journal: Materials Today Bio

Article Title: CRISPR-engineered human lung organoids with a biomolecular condensate reporter enable mechanistic toxicity monitoring

doi: 10.1016/j.mtbio.2026.102972

Figure Lengend Snippet: Dose–response profiling of representative toxicant groups in A549 and G3BP1–mCherry hLO. Cell viability was measured after 24 h exposure to five representative toxicants in two in vitro lung models: A549 (2D monolayer) and G3BP1–mCherry knock-in human lung organoids (hLOs). (A) Dose–response curves for the disinfectant/biocide group— didecyldimethylammonium chloride (DDAC), 2-octyl-4-isothiazolin-3-one (OIT), and polyhexamethylene guanidine (PHMG-p). (B) Dose–response curves for industrial chemicals—4,4′-MDI and acrylonitrile. Curves were fitted using nonlinear regression (log[inhibitor] vs . normalized response, variable-slope model) in GraphPad Prism. Data are presented as mean ± SD. (C) Heatmap displaying relative log 2 (IC 50 ) values comparing A549 and hLO models. Each IC 50 value was normalized to the mean IC 50 of the respective compound. The color scale represents deviations from the mean, with blue indicating higher sensitivity and red indicating lower sensitivity. The scale ranges from −2 to +2, corresponding to up to a fourfold deviation from the mean IC 50 ; values outside this range were clipped for clarity.

Article Snippet: A549 human lung epithelial carcinoma cells, purchased from the American Type Culture Collection (Manassas, VA, USA), were cultured in Dulbecco's modified eagle medium, high glucose (LM001-05, WELGENE, Gyeongsan, Korea) supplemented with 10% fetal bovine serum (16000044, Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (15040122, Gibco), 1 mM sodium pyruvate (11360070, Gibco), and 1 × GlutaMAX (35050061, Gibco).

Techniques: In Vitro, Knock-In

Journal: Materials Today Bio

Article Title: CRISPR-engineered human lung organoids with a biomolecular condensate reporter enable mechanistic toxicity monitoring

doi: 10.1016/j.mtbio.2026.102972

Figure Lengend Snippet: Comparative toxicant sensitivity and stress granule responses in G3BP1–mCherry hLOs and conventional lung models. (A) Cell viability analysis of sodium arsenite (NaAsO 2 ), benzalkonium chloride (BAC), and 1,2-benzisothiazolin-3-one (BIT) was evaluated in three lung models: A549 (3D spheroid), patient-derived lung cancer organoid (LCO), and G3BP1–mCherry knock-in lung organoid (hLO). IC 50 values were determined by nonlinear regression (log[inhibitor] vs . normalized response, variable-slope model) using GraphPad Prism. Data are presented as mean ± SD. (B) Heatmap depicting relative log 2 (IC 50 ) values to illustrate model-specific differences in toxicant sensitivity. Each IC 50 value was normalized to the mean IC 50 of the respective compound. Blue and red represent higher and lower sensitivity relative to the mean, respectively. (C) Immunofluorescence staining showing stress granule (SG) formation in patient-derived LCOs and G3BP1–mCherry hLOs after 1 h exposure to sodium arsenite (0, 50, or 100 μM). G3BP1 (green) indicates SG localization, and nuclei were counterstained with Hoechst 33342 (blue). Representative images at intermediate concentrations (12.5, 25, 75 μM) are shown in . Scale bars, 20 μm. (D) Quantification of SG numbers per cell in patient-derived LCOs and hLOs following sodium arsenite treatment (0–100 μM). Bars represent mean ± SD. Statistical analysis was performed using two-way ANOVA followed by Sidak's multiple comparisons test (ns, not significant; *** p < 0.001; **** p < 0.0001).

Article Snippet: A549 human lung epithelial carcinoma cells, purchased from the American Type Culture Collection (Manassas, VA, USA), were cultured in Dulbecco's modified eagle medium, high glucose (LM001-05, WELGENE, Gyeongsan, Korea) supplemented with 10% fetal bovine serum (16000044, Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (15040122, Gibco), 1 mM sodium pyruvate (11360070, Gibco), and 1 × GlutaMAX (35050061, Gibco).

Techniques: Derivative Assay, Knock-In, Immunofluorescence, Staining