hPSC-derived microglia shape neuronal morphology and enhance network activity in vitro 163 5 Figure 3: Microglia have significant effect on activity of neuronal co-cultures. A) Schematic representation of spike, burst and network burst events. B) Representative example of 100 s of baseline activity at 12 weeks post-plating of co-cultures without (-MG) and with (+MG). C) Quantification of the activity shows increased number of active channels (P = 0.010) and D) increased spike frequency (P = 0.030) in cultures with microglia. E) Burst Rate and F) Network Burst Rate show a similar but non-significant trend of increased activity. N = 12 MEA well observations on 4 neuronal backgrounds. WPP = weeks post plating (of neurons). Shaded error bars represent the standard deviation. *P < 0.05. ALD microglia-like cells suitable for co-culture Now that we established an in vitro model for hPSC-derived microglia co-culture, we wanted to explore whether this model can be applied in a disease relevant context. As mentioned before, in ALD a primary involvement of microglia in the cerebral demyelination phenotype is suspected. Hence, we generated microglia-like cells of 8 ALD cell lines, and 4 controls. We applied the same co-culture procedure as in the previous experiments (Fig. 1A) but instead of plating the neurons in separate wells, a mixture of all batches was equally divided over all wells.
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