Electrophysiological characterization of NSCs after differentiation induced by OEC conditioned medium.
ABSTRACT We induced neural stem cells (NSCs) to neurons by olfactory ensheathing cell (OEC) conditioned medium and characterized their electrophysiological properties after neuronal differentiation.
Fetal NSCs and OECs were cultured from embryonic day 14 SD rats and the conditioned medium was collected and stored at -20°C when the cell number was up to 80% of the culture flasks. The experiment groups were divided into a control group (cultured with DMEM/F12 without FBS) and an OECs induction group (cultured with OEC conditioned medium and DMEM/F12 without FBS). Immunocytochemistry staining was carried out to identify the neurons derived from NSCs and their electrophysiological properties were characterized after neuronal differentiation using a patch-clamp technique.
The NSCs divided rapidly in the expansion medium, forming small proliferating spheres after 7 days. The OECs induction group presented an evident neuron-like type 7 days after adding OEC conditioned medium, and the nestin immunochemistry staining was positive. The electrophysiological characterization showed that the derived neurons presented a transient inward sodium current and slow outward potassium current under proper electric stimulus, which were blocked by tetrodotoxin (TTX) and tetraethylammonium (TEA).
OEC conditioned medium can induce NSCs to form neurons, and electrophysiological characterization demonstrated that the derived neurons presented active electrophysiological properties which are essential for nervous excitation.