Fig. 4

BrainPhys medium + B27/GFs supports neuronal activity comparable to NBM. (A) Schematic overview of the differentiation protocol used for BrainPhys medium. Neurons were plated in half DMEM/half BrainPhys with or without supplementation. At DIV10, media was gradually changed to 100% BrainPhys. (B) Representative images of jRGECO signal in cultures with BP with and without supplementation at WIC6. Scale bar, 100 μm. (C) Example traces of jRGECO intensity of cultures in BP with or without supplementation. Grey lines show traces of individual cells and red lines show the average signal in the FOV. (D) Percentage silent cells in a FOV. N numbers of individual experiments and number of FOVs in brackets. B/G/C: 7(37); BP + B/G/C: 7(15); BP -GF: 4(1); BP + B/G/NT3: 7(2); BP + B/G/C + B27: 4(24); BP + B/G/NT3 + B27: 3(18). (E) The mean frequency of calcium events of individual neurons in a FOV. B/G/C: 7(38); BP + B/G/C: 7(39); BP -GF: 4(21); BP + B/G/NT3: 7(39); BP + B/G/C + B27: 4(24); BP + B/G/NT3 + B27: 3(18). (F) The mean frequency of network bursts in a FOV. (G) Percentage of synchronicity, defined by the percentage cells active at the same time. Bar plots represent the mean ± SEM. Each dot represents an a FOV. Kruskal-Wallis H test with Dunn’s correction: * P < 0.05, ** P < 0.01, *** P < 0.001. ns = non-significant, P > 0.05.