Figure 4.

NPCs differentiated in remodelable hydrogels mature into neurotransmitter‐responsive neurons. After 1 week of remodeling followed by 2 weeks of differentiation, encapsulated NPCs were incubated with the calcium‐sensitive Fluo‐4AM dye prior to treatment with either A) γ‐aminobutyric acid (GABA) or B) glutamate. Time lapse confocal microscopy was used to image changes in intracellular calcium concentration. Examples of different types of cellular responses to neurotransmitter treatment are presented. To confirm the imaged cells were differentiated neurons, after calcium imaging, the samples were fixed and immunostained for the differentiated neuronal markers β‐tubulin III and microtubule‐associated protein 2 (MAP2).