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. 2022 Jun 2;18(2):404–409. doi: 10.4103/1673-5374.346466

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Copyright: © Neural Regeneration Research

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BDNF-hNSC-Exo promotes the survival and differentiation of neural stem cells in vitro.

(A) The expression level of BDNF was measured by western blotting after the addition of BDNF-hNSC-Exo to cultured neural stem cells. Data were normalized to β-actin expression. (B) The differentiation efficiency of BDNF-hNSC-Exo was evaluated using quantitative reverse transcription-polymerase chain reaction. Data were normalized to GAPDH mRNA expression. Cells were treated with a 500 μM solution H₂O₂ for 5 hours. (C) Western blot analysis of Tuj1 in neural stem cells. Cells were treated with a 500 μM solution H₂O₂ for 5 hours. Data were normalized to GAPDH expression. (D) Immunofluorescence showed that the ratio of Tuj1-positive cells (red) in the BDNF-hNSC-Exo group was higher than that in the control group. The percentage of Tuj1-positive cells was obtained by counting cells in three visual fields. Cells were treated with a 500 μM solution H₂O₂ for 5 hours. Nuclei were stained by DAPI (blue). Scale bars: 50 μm. (E) BDNF-hNSC-Exo remarkably increased cell survival in the H₂O₂ stress model (CCK8 assay). Cells were treated with a 500 μM solution H₂O₂ for 5 hours. (F) TUNEL-positive cells. The proportion of TUNEL-positive cells in the BDNF-hNSC-Exo group was remarkably lower than that in the PBS group. Cells were treated with a 500 μM solution H₂O₂ for 5 hours. Nuclei were stained by DAPI (blue). Scale bars: 50 μm. (G) Western blotting showed that the expressions of apoptosis-related proteins cleaved-caspase-3 and Bax were significantly decreased in the BDNF-hNSC-Exo group compared with the other two groups. Cells were treated with a 500 μM solution H₂O₂ for 5 hours. Data were normalized to GAPDH expression. All experiments were conducted in triplicate. All data are presented as mean ± SEM. *P < 0.05 (one-way analysis of variance followed by Tukey’s post hoc test). Bax: Bcl2 associated X, apoptosis regulator; BDNF: brain-derived neurotrophic factor; Exo: exosomes; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hNSC: human neural stem cell; Tuj1: β-tubulin III.