Fig 2: Antioxidant treatment with superoxide dismutase (SOD) preserves telomere length, reduces oxidative stress and senescence, and improves angiogenic balance in LOPE trophoblast organoids.a, Workflow for sample collection, trophoblast stem cell isolation, organoid formation and antioxidant treatment with superoxide dismutase (SOD). b, Schematic of SOD catalysing the dismutation of superoxide anions into oxygen. c, Telomere length dynamics over 4 weeks in culture (left) and violin plots of telomere length at endpoint (right) in control and LOPE organoids with or without SOD treatment. d, Representative immunofluorescence images of control and LOPE trophoblast organoids stained for γH2AX (DNA damage, green), 8-OHdG (oxidative stress, magenta) and DAPI (nuclei, blue) in the presence or absence of SOD. Scale bar, 100 μm. e, Quantification of 8-OHdG and f, γH2AX fluorescence intensity in control and LOPE trophoblast organoids with or without SOD treatment. Data presented as Tukey’s box and whisker plots. g, Representative immunohistochemistry images showing SA-β-Gal (violet) to assess senescence, co-stained with pan-cytokeratin to image trophoblast cells and counterstained with DAPI for nuclei; imaged at 40× magnification, and h, quantification of SA-β-Gal-positive staining. i-k, Concentrations of angiogenic factors in culture media after 4 weeks, including i, sFlt-1 (pg/mL), j, PlGF (pg/mL), and k, sFlt-1/PlGF ratio, in control and LOPE trophoblast organoids with or without SOD treatment. Data are presented as mean ± s.d. Trophoblast organoids were generated from n = 3 separate biological samples per fetal sex, per group; experiments performed in technical triplicate. For c,e-f,h,i-k, statistical analysis was performed using Welch’s ANOVA with multiple comparisons. ****p < 0.0001; ns, not significant. Source data file is provided.