Fig. 6

Nutrient utilization and metabolic profile during late-stage (S5D3–S7D8) differentiation toward hESC-derived insulin-producing cells indicates the cells rely primarily on oxidative phosphorylation similar to donor human islets. A C-peptide, B proinsulin and C C-peptide to proinsulin ratio measured in spent media samples. For panels A-C, the solid line = mean (n = 4–9 biological replicates). D Intracellular insulin content (n = 6–11 biological replicates; mixed-effects analysis with Tukey post-test). E Glucose concentration with percent glucose consumed (n = 9–17 biological replicates, 5 donor human islets; orange dash line indicates glucose concentration in fresh media; mixed-effects analysis with Tukey post-test). F Lactate concentration measured in stage-specific spent media (n = 15–20 biological replicates, 5 donor human islet preps; orange dash line indicates lactate concentration in fresh media; mixed-effects analysis with Tukey post-test). For panels A-F, # indicates the spent media were sampled 48 ± 2 h from the previous media change; otherwise, spent media were sampled 24 ± 2 h from the previous media change. G Glucose consumption rates, H lactate production rates (G-H n = 4–13 biological replicates, 5 donor human islets; mixed-effects analysis with Tukey post-test). I lactate yield per glucose consumed in stage-specific spent media (n = 12–15 biological replicates, 5 donor human islet preps; mixed-effects analysis with Tukey post-test). J Oxygen consumption rates (n = 3 biological replicates and 3 donor human islets). All box and whisker plots show individual points with median and interquartile range; bar charts show individual points with mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Spent media were sampled 24 ± 2 h from the previous media change unless indicated otherwise; S5D3 = stage 5 day 3, S6D7 = stage 6 day 7, S7D4 = stage 7 day 4, S7D8 = stage 7 day 8, S7D12 = stage 7 day 12