Research Paper Volume 13, Issue 1 pp 262—278

Combined transplantation of neural stem cells and bone marrow mesenchymal stem cells promotes neuronal cell survival to alleviate brain damage after cardiac arrest via microRNA-133b incorporated in extracellular vesicles

Combined treatment of BMSCs and NSCs promotes survival of neuronal cells in vivo and in vitro. Neuronal cells injured by hypoxia were untreated (H) or co-cultured with BMSCs (H-BMSCs), NSCs (H-NSCs) or BMSCs + NSCs (H-Co) in Panels (A and B). The sham-operated rats were not treated with any cells (sham) while CA rats were not treated (Model) or injected with BMSCs, NSCs or BMSCs + NSCs in Panels (C–J). (A) Apoptosis of neuronal cells assessed by TUNEL staining (× 200). (B) Cell viability assessed by CCK-8 assay. (C) Representative images of the NeuN-positive cells in the hippocampal CA1 region visualized using immunofluorescence staining (× 400). (D) Representative images of NeuN-positive cells in the cerebral cortex visualized using immunofluorescence staining (× 400). (E) The number of NeuN-positive cells in the hippocampal CA1 region. (F) The number of NeuN-positive cells in the cerebral cortex. (G) Apoptosis of neuronal cells in the hippocampal CA1 region assessed by TUNEL staining (× 400). (H) Apoptosis of neuronal cells in the cerebral cortex assessed by TUNEL staining (× 400). (I) Comparison of apoptotic rate in the hippocampal CA1 region. (J) Comparison of apoptotic rate in the cerebral cortex. * p vs. the Control (neuronal cells without any treatment) or Model (rats with CA without any treatment) group, # p vs. the H group (hypoxia-induced injured neuronal cells without any treatment). Data were expressed as mean ± standard deviation, and comparison among multiple groups were analyzed by one-way ANOVA followed by Tukey's post hoc test. n = 10 in animal experiments. The cell experiments were conducted 3 times independently.

Figure 2. Combined treatment of BMSCs and NSCs promotes survival of neuronal cells in vivo and in vitro. Neuronal cells injured by hypoxia were untreated (H) or co-cultured with BMSCs (H-BMSCs), NSCs (H-NSCs) or BMSCs + NSCs (H-Co) in Panels (A and B). The sham-operated rats were not treated with any cells (sham) while CA rats were not treated (Model) or injected with BMSCs, NSCs or BMSCs + NSCs in Panels (CJ). (A) Apoptosis of neuronal cells assessed by TUNEL staining (× 200). (B) Cell viability assessed by CCK-8 assay. (C) Representative images of the NeuN-positive cells in the hippocampal CA1 region visualized using immunofluorescence staining (× 400). (D) Representative images of NeuN-positive cells in the cerebral cortex visualized using immunofluorescence staining (× 400). (E) The number of NeuN-positive cells in the hippocampal CA1 region. (F) The number of NeuN-positive cells in the cerebral cortex. (G) Apoptosis of neuronal cells in the hippocampal CA1 region assessed by TUNEL staining (× 400). (H) Apoptosis of neuronal cells in the cerebral cortex assessed by TUNEL staining (× 400). (I) Comparison of apoptotic rate in the hippocampal CA1 region. (J) Comparison of apoptotic rate in the cerebral cortex. * p < 0.05 vs. the Control (neuronal cells without any treatment) or Model (rats with CA without any treatment) group, # p < 0.05 vs. the H group (hypoxia-induced injured neuronal cells without any treatment). Data were expressed as mean ± standard deviation, and comparison among multiple groups were analyzed by one-way ANOVA followed by Tukey's post hoc test. n = 10 in animal experiments. The cell experiments were conducted 3 times independently.