Fig. 3

Nutrient utilization and metabolic profile during differentiation toward pancreatic progenitors. A Schematic of cell morphology (2D vs 3D) during four-stage differentiation. B Glucose concentrations (n = 7–14 biological replicates, one-way ANOVA with Tukey post-test; orange dash line indicates glucose concentration in fresh media) and C lactate concentration measured in spent media (n = 7–14 biological replicates, one-way ANOVA with Tukey post-test; orange dash line indicates lactate concentration in fresh media). D Glucose consumption rates (n = 5–7 biological replicates; one-way ANOVA with Tukey post-test) and E lactate production of S1D3 to S4D4 cells (n = 8–10 biological replicates; one-way ANOVA with Tukey post-test). F Glucose concentration and G lactate concentration in the spent media between S1D1 and S1D2 (n = 3 biological replicates); panels F-G: media were samples at 2.5, 4.5, 6.5 and 24 h after the last media change; orange dots represent concentration in fresh media. H Lactate yield per glucose consumed in stage-specific spent media (n = 6–13 biological replicates, one-way ANOVA with Tukey post-test). I Basal OCR and J Area under the curve (AUC) of stem cell derivatives, undifferentiated H1 cells and MIN6 cells (n = 3—4 biological replicates, one-way ANOVA with Tukey post-test). 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 stated otherwise; the x-axis labels on all panels denote the stage and day, e.g., S1D1 = stage 1 day 1