Supplementary Figure 8: Co-culture of hPSC-derived neurons with astrocyte or astrocyte conditioned medium.

(a) Schematic illustration of long-term maintenance of P8S10D+D neurons with astrocyte co-culture or conditioned media. (b) Bright field images of P8S10D+D neurons co-cultured with astrocytes or conditioned media at day 25 and day 35. (c) Representative traces of action potential firings of P8S10D+D neurons co-cultured with astrocytes or conditioned media at day 25 and day 35, evoked by current injection from -30 to +100 pA. (d) Bright field images of P8S10D+D neurons co-cultured with astrocytes at day 90. N = 15, 23, 7, 5 cells recorded for cultures with astrocytes at day 25, 35, and cultures in conditioned medium at day 25, 35. (e) Quantitative analysis of passive membrane properties and action potential properties. (f) MAP2ab staining of P8S10D+D neurons with astrocytes co-culture shows increased complexity of dendrite branching with time in culture. (g) Sholl analysis at day 36 of P8S10D+D neurons co-cultured with astrocytes, compared with neurons cultured with conditioned medium alone. Scale bars: 50 μm. Error bars represent s. e. m. *** P<0.001.