Fig. 3: EMS medium alone is not sufficient to induce islet dysfunction. NF and EMS media maintain islet viability and functionality including glucose stimulated insulin response for 8 days in the standalone PANIS.

A Representative live/dead assay image for islets cultured in NF and EMS media. B Percentage of live cell shows no significant difference between NF and EMS (n = 10 islets and n = 7 islets). C Immunofluorescence labeling for C-peptide expressing β-cells and glucagon expressing α-cells for islets cultured in NF and EMS. D The fraction of quantified C-peptide and glucagon positive cells shows no significant difference (n = 8 islets and n = 10 islets). E Representative images showing mitochondrial associated ROS immunofluorescence labeling of islets. F Percentage of ROS intensity quantification with respect to a positive control (50 µM TBHP) for islets cultured in NF and EMS media (see Methods) demonstrates no significant difference between NF and EMS (n = 8 islets and n = 8 islets). G Standalone PANIS efflux analysis showed the change in islet-derived factors in EMS normalized to NF (see Fig. S6), with no significant difference between them (n = 5 MPS). H Glucose stimulated insulin secretion (GSIS) assay, where islet samples were subjected to low (3 mM), and subsequent high (16 mM) glucose concentration and the resulting secreted insulin was measured and showed no significant difference at either glucose concentration in islets that were maintained in either NF or EMS media (n = 4 islets and n = 3 islets). I Comparison of insulin stimulation index secretion for islets cultured in NF and EMS showing no statistically significant difference. Scatter plot graphs display the mean and standard error (SEM) with p-values shown. For graphs -B, D, F, H, and I- statistical analysis was done by two-tail t test with Welch’s correction. For graph -G- Statistical analysis was done by Wilcoxon signed-rank test of normalized EMS values to their corresponding NF study. All scale bar, 50 µm.