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hNSCs characterization in vitro . (A) Morphology of hNSCs (the four ball structures with clear edges are the neurospheres aggregated by hNSCs). Photomicrograph was taken from culture at 7 days after neurospheres formed (scale bar: 100 µm). (B, C) Flow cytometry showed high levels of NSC markers <t>(CD133,</t> 91.4%; nestin, 98.7%; and Sox2, 98.83%). (D–F) Immunofluorescence images showed that the induced hNSCs were positive for GFAP (green), Tuj-1 (red) and O4 (red). Nuclear staining (DAPI, blue) is included in merged panel. Scale bars: 50 µm. (G) Bar graph showing the average percentage of cells positive for neural stem cell markers CD133, nestin, and Sox2 in flow cytometry plots (B, C). (H) Bar graph showing the average percentage of hNSCs expressing the three lineages in the immunofluorescence images (D–F): astrocytes (GFAP), neurons (Tuj-1), and oligodendrocytes (O4). Data are presented as the mean ± standard error of the mean (SEM). DAPI: 4’,6-Diamidino-2-phenylindole; GFAP: glial fibrillary acidic protein; hNSCs: human neural stem cells.
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hNSCs characterization in vitro . (A) Morphology of hNSCs (the four ball structures with clear edges are the neurospheres aggregated by hNSCs). Photomicrograph was taken from culture at 7 days after neurospheres formed (scale bar: 100 µm). (B, C) Flow cytometry showed high levels of NSC markers (CD133, 91.4%; nestin, 98.7%; and Sox2, 98.83%). (D–F) Immunofluorescence images showed that the induced hNSCs were positive for GFAP (green), Tuj-1 (red) and O4 (red). Nuclear staining (DAPI, blue) is included in merged panel. Scale bars: 50 µm. (G) Bar graph showing the average percentage of cells positive for neural stem cell markers CD133, nestin, and Sox2 in flow cytometry plots (B, C). (H) Bar graph showing the average percentage of hNSCs expressing the three lineages in the immunofluorescence images (D–F): astrocytes (GFAP), neurons (Tuj-1), and oligodendrocytes (O4). Data are presented as the mean ± standard error of the mean (SEM). DAPI: 4’,6-Diamidino-2-phenylindole; GFAP: glial fibrillary acidic protein; hNSCs: human neural stem cells.

Journal: Neural Regeneration Research

Article Title: Transplantation of human neural stem cells repairs neural circuits and restores neurological function in the stroke-injured brain

doi: 10.4103/NRR.NRR-D-24-00363

Figure Lengend Snippet: hNSCs characterization in vitro . (A) Morphology of hNSCs (the four ball structures with clear edges are the neurospheres aggregated by hNSCs). Photomicrograph was taken from culture at 7 days after neurospheres formed (scale bar: 100 µm). (B, C) Flow cytometry showed high levels of NSC markers (CD133, 91.4%; nestin, 98.7%; and Sox2, 98.83%). (D–F) Immunofluorescence images showed that the induced hNSCs were positive for GFAP (green), Tuj-1 (red) and O4 (red). Nuclear staining (DAPI, blue) is included in merged panel. Scale bars: 50 µm. (G) Bar graph showing the average percentage of cells positive for neural stem cell markers CD133, nestin, and Sox2 in flow cytometry plots (B, C). (H) Bar graph showing the average percentage of hNSCs expressing the three lineages in the immunofluorescence images (D–F): astrocytes (GFAP), neurons (Tuj-1), and oligodendrocytes (O4). Data are presented as the mean ± standard error of the mean (SEM). DAPI: 4’,6-Diamidino-2-phenylindole; GFAP: glial fibrillary acidic protein; hNSCs: human neural stem cells.

Article Snippet: Cells were then characterized by flow cytometry (CytoFlex, Beckman Coulter, CA, USA, AS51394) using the NSC markers nestin (mouse monoclonal antibody, BD Biosciences, San Jose, CA, USA, Cat# 561230, RRID: AB_10562398), Sox2 (mouse monoclonal antibody, BD Biosciences, Cat# 561610, RRID: AB_10712763), and CD133 (mouse monoclonal antibody, Miltenyi Biotech, Bergish Gladbach, Germany, Cat# 130-113-186, RRID: AB_2726012).

Techniques: In Vitro, Flow Cytometry, Immunofluorescence, Staining, Expressing