Chunk #7 — Results — Human neuron/astrocyte co-culture in combination with simultaneous differentiation of NPCs improves synchronization of calcium oscillations and network activity
In order to compare fully human co-cultures with mixed rodent/human co-cultures (as previously described2931), we cultured our hiPSC-derived neurons with freshly dissected rat primary astrocytes in the presence of DAPT. Mixed rat/human co-cultures showed an increased percentage of active neurons, bursting frequency and synchronization of neuronal calcium oscillations (Supplementary Fig. S2) 2 to 5 weeks after final plating compared to monocultures, which is much faster than in previous studies (>2 months)2931. Additionally, there is a significantly increased percentage of active neurons, bursting frequency and correlation score in mixed co-cultures compared to fully human co-cultures at the earliest time points (wk 2 and 3), while the percentage of active neurons decreased significantly at the latest time point measured (wk5). Nevertheless, with our aim to generate a fully human model, we only focus on human astrocytes for all further co-culture experiments.
