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. 2020 Apr 30;21(9):3191. doi: 10.3390/ijms21093191

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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LS neural stem cells (NSCs) manifested similar proliferative and differentiation capacity. (A) Immunofluorescence analysis of the neural stem cell marker nestin, manifesting an efficient generation of NSCs from iPSCs; scale bar: 100 µm. (B) Electropherograms showing the mutation m.13513G>A in patient NSCs and its absence in control NSCs (left) and heteroplasmy levels of m.13513G>A mutation by RFLP followed by Agilent quantification. (C) Proliferation assay of NSCs with the thymidine analogue 5-ethynyl-2’-deoxyuridine (EdU). Scale bar: 15 µm. (D) Quantification of EdU (percentage of EdU+/Hoechst+). (E) Bright field images (4× and 10×) of neural populations obtained after differentiation of NSCs. (F) Immunofluorescence analysis of MAP2, a marker of mature neurons, and GFAP, a marker of astrocytes, in the neural populations obtained after differentiation of NSCs in N2B27 for 3 weeks (G–H) Immunofluorescence analysis of the GABAergic marker GAD 65/67 and glutamatergic marker KGA together with neuronal markers (Tuj1 and MAP2).