Figure 4.

Electrophysiological and pharmacological profiling of SNs derived from human iPSCs

Cells were replated on multi-electrode array (MEA) plates on day 11.

(A) Raster plot of spontaneous activity of one G3_Hybrid culture (one well of a 96-well plate containing 8 electrodes). Each row represents the response from a single electrode, where each vertical line represents a spike (supra-threshold extracellular potential). The scale bar shows 100 ms.

(B) The average spike rate of cells in each well (each dot represents one well). Plots show median spike rates for each time point, and error bars show SD.

(C) Cultures were exposed to a panel of agonists to assess the presence of functional TRP channels. Plots present the average spike rate for individual wells before, during, and after exposure to noxious stimuli. Each dot represents the average spike rate for one culture; bars show means for each treatment group, and error bars show SD. ∗adjusted p < 0.05, ∗∗adjusted p < 0.01, ∗∗∗adjusted p < 0.001 as derived from Sidak’s multiple comparisons tests after 2-way repeated-measures ANOVA.

(D) Spike shapes recorded for each electrode of a representative culture, where the average spike rate for each is shown above the trace. Horizontal lines in each show the spike threshold for each electrode.

(E) The response of single units before and during treatments was analyzed (G3_H condition). The main plot shows the cumulative probability of firing rates across the conditions, where KCl represents a QC treatment to identify responsive cells. Inset: violin plots of the responses, where each dot shows the firing rate of a single neuron unit.

(F) The percentage of single units that responded to each of the treatments. Data show means and error bars show SD. ∗adjusted p < 0.05 and ∗∗adjusted p < 0.01 as determined from Tukeys’s multiple comparisons tests after two-way ANOVA.

Data are the result of 267 individual cultures across three independent differentiations.