Fig. 5. Long-term culture of CN-differentiated cells show bipotential in response to different environmental niche. (a~c) Most of the cells matured into astrocytes in DMEM-FBS. No neuronal activity was observed in those astrocytes. (d~f) Fully matured neuronal cells (MAP2+; Red /NeuN+; Green) were observed in ITSFn media, and those neuronal cells showed high level of Ca2+ transient in response to high KCl. (g) Ca2+ response of cells in ITSFn media by 30 mM K+ in the absence or presence of 2 M nimodipine. (h) Whole cell recordings of CN-differentiated cells under voltage clamp held at -70 mV. Voltage steps of 10 mV were applied to the cell under voltage clamp. Each trace indicated a recording of different cells. (i) CN-derived neuroblasts matured into neuron for 4 weeks analyzed by current injection of 20 pA steps under current clamp. Action potentials were blocked by 0.5 uM TTX. Out of 14 cells, 3 cells showed action potentials. After TTX was washed off, action potentials were revived. (j) The biocytin solution was filled into the electrodes, and injected to the cells which show action potectial for morphological analysis, and these cells these cells were confirmed as MAP2+ by immunostaining. Scale bars, 50 µm (b, e, j).
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