2D- and 3D-cultures of human trabecular meshwork cells: A preliminary assessment of an in vitro model for glaucoma study
Fig 1
2D- and 3D-HTMC oxidative stress sensitivity.
Panel A- Confocal microscopy analyses of nucleus and cytoskeletal markers were performed on untreated and H2O2-treated 2D- and 3D HTMC cells after 72 hrs of experimental procedures. Representative images are related to immunoreactivity for To-ProTM and Phalloidin, as nuclear and cytoskeleton markers, respectively. Merged images showed cytoskeleton plus Nucleus. Fluorescence signals were captured at 60x magnification. Panel B- DCF assay. DCF assay was performed on untreated and H2O2 (U.T.)-treated 2D- and 3D HTMC cells and the fluorescence was recorded at 1, 2 and 4 hrs of experimental procedures. Data are expressed as % of ROS production in untreated HTMCs after 1hr and represent the mean ± SD of 3 independent experiments, each performed six times. White and grey bars = 2D and 3D cultures, respectively ***p <0.001 vs respective untreated cultures (1h); §§§p<0.001 3D-permanent H2O2 vs 3D-pulse H2O2; @@@p<0.001 3D permanent H2O2 vs 2D permanent H2O2; ++p<0.01, +++p<0.001 3D pulse H2O2 vs 2D pulse H2O2 (Two-way ANOVA followed by Bonferroni posttest).