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

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Stiff substrate promotes A549 and H460 cell migration and down-regulates Piezo1 channel e xpression. (A–D) Transwell assay of the effects of substrate stiffness on cell migration. Representative images of migrated cells stained with crystal violet (10x, A-B) and statistical analysis of data from three independent experiments (C–D). Scale bar: 50 μm. (E–H) Flow cytometry assessing the effects of substrate stiffness on cell surface Piezo1 protein expression. Representative images of flow cytometry (E–F) and statistical analysis of data from three (G–F) independent experiments. All data were normalized to that of 3 kPa group. Data were presented as mean ± SD. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗ P < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Two human non-small lung cancer cell lines, A549 and H460, were used in the study and they were obtained from the American Type Culture Collection (ATCC) and cultured in 1640 medium (#11875119, Gibco, USA) containing 10 % fetal bovine serum (FBS, #11011–8611, Sijiqing, China), 100 U/ml penicillin and 100 μg/ml streptomycin (#15140122, Gibco, USA) at 37 °C in a 5 % CO 2 incubator.

Techniques: Migration, Transwell Assay, Staining, Flow Cytometry, Expressing

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Piezo1 channel negatively regulates substrate stiffness-induced A549 cell migration. (A, D) Piezo1 channel blockade with GsMTx4 promotes cell migration on both soft and stiff substrates. (B, E) Piezo1 channel activation with Yoda 1 inhibits cell migration on both soft and stiff substrates. (C, F) Piezo1 channel knockdown with specific siRNA transfection promotes cell migration on both soft and stiff substrates. Representative images of migrated cells stained with crystal violet (10x, A-C) and statistical analysis of data from three independent experiments (D–F). Scale bar: 50 μm. All data were normalized to the 3 kPa group. Data were presented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Two human non-small lung cancer cell lines, A549 and H460, were used in the study and they were obtained from the American Type Culture Collection (ATCC) and cultured in 1640 medium (#11875119, Gibco, USA) containing 10 % fetal bovine serum (FBS, #11011–8611, Sijiqing, China), 100 U/ml penicillin and 100 μg/ml streptomycin (#15140122, Gibco, USA) at 37 °C in a 5 % CO 2 incubator.

Techniques: Migration, Activation Assay, Knockdown, Transfection, Staining

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Filopodia formation in A549 cells on 3, 10 and 20 kPa substrates, respectively. Representative images of filopodia morphology (A) and statistical analysis of the filopodia length (B) and number (C) from indicated number of cells. Red, F-actin staining with rhodamine-labeled phalloidin; blue, nucleus staining with Hoechst 33342. All data were normalized to that of the 3 kPa group. Scale bar: 20 μm. Data were presented as mean ± SD. ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Two human non-small lung cancer cell lines, A549 and H460, were used in the study and they were obtained from the American Type Culture Collection (ATCC) and cultured in 1640 medium (#11875119, Gibco, USA) containing 10 % fetal bovine serum (FBS, #11011–8611, Sijiqing, China), 100 U/ml penicillin and 100 μg/ml streptomycin (#15140122, Gibco, USA) at 37 °C in a 5 % CO 2 incubator.

Techniques: Staining, Labeling

Journal: Materials Today Bio

Article Title: Photocrosslinkable lung dECM hydrogels promote stiffness-dependent lung cancer growth and chemoresistance

doi: 10.1016/j.mtbio.2026.102838

Figure Lengend Snippet: NSCLC growth is dependent on matrix type and stiffness. (A) Viability staining of encapsulated A549 cells using fluorescein diacetate (FDA, green, live) and propidium iodide (PI, red, dead). Day 1 scale = 200 μm, day 7 and 14 scales = 500 200 μm. (B) Representative fluorescent images of A549-laden cultures fixed on days 3, 7, and 14 and stained with DAPI (blue, nuclei) and phalloidin (red, actin) to observe cell morphology and distribution over time. Scale = 250 μm, inset scale = 100 μm. (C) Cell viability of A549s within matrices measured through live-dead quantification of FDA or PI stained live (green) and dead (red) cells over time via ImageJ analysis. N = 1. n = 3. (D) Metabolic activity of A549 cells within matrices was determined by PrestoBlue metabolic assay. N = 1. n = 6. (E) DNA content of A549 cells within matrices was determined by PicoGreen™ DNA quantification. N = 1. n = 6. (F) Number of nuclei per ROI was determined via Image J analysis of DAPI-stained cells within ROIs captured using an Evident FV4000 confocal microscope at 10× magnification. N = 2. n = 4–8 ROIs were captured per condition per timepoint. (G) Area of actin per ROI (μm 2 ) was determined through ImageJ analysis of phalloidin-stained cell actin fibers. N = 2. n = 4–8. All data is represented as mean ± SD.

Article Snippet: 1 × 10 6 A549 cells (ATCC) were seeded at P90 in T75 culture flasks and cultured using 10 mL complete A549 media (Dulbecco's modified eagle medium (DMEM) + 10 % FCS + 1 % pen/strep) in a tissue culture incubator at 37 °C.

Techniques: Staining, Activity Assay, Metabolic Assay, Microscopy