Fig. 3.

Effect of different glucose levels on soma size and proliferation during neuronal differentiation. (A) Flow cytometry signals of the forward scatter (FS Lin) as a measure of cell size for TSC2 +/+ (red) and TSC2 −/− (blue) cultures during Livesey differentiation at timepoints days 10, 18, 50, and 80 in high and low glucose conditions (n = 1 for each differentiation). No differences in cell size between the genetic conditions could be seen in 25 mM glucose media. However, soma size differed in 5 mM glucose media at the NSC (day 18) and neuronal (day 80) stage with TSC2 −/− cells being smaller at day 18 but larger at day 80 than TSC2 +/+ cells. (B) Growth curves obtained from TSC2 +/+ and TSC2 −/− NSCs. Cells were cultured for 3–4 days in 25 mM culture media or 5 mM culture media (n = 9 for each condition). In high glucose media, TSC2 −/− NSCs doubled in a similar time to TSC2 +/+ NSCs (TSC2 +/+: 18.8 h, TSC2 −/−: 16.7 h). No differences in growth curve and doubling time could be observed in TSC2 −/− NSCs when transferred to 5 mM glucose media (20.3 h), whereas a reduction in glucose concentration completely abolished proliferation of TSC2 +/+ NSCs. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)