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  • Title: Electrophysiological characterization of NSCs after differentiation induced by OEC conditioned medium.
    Author: Duan D, Rong M, Zeng Y, Teng X, Zhao Z, Liu B, Tao X, Zhou R, Fan M, Peng C, Chen P, Liang S, Lu M.
    Journal: Acta Neurochir (Wien); 2011 Oct; 153(10):2085-90. PubMed ID: 21301894.
    Abstract:
    PURPOSE: We induced neural stem cells (NSCs) to neurons by olfactory ensheathing cell (OEC) conditioned medium and characterized their electrophysiological properties after neuronal differentiation. METHODS: 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. RESULTS: 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). CONCLUSION: 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.
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