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

Neuronal cell survival in rats with CA is promoted by miR-133b transferred via BMSCs-derived EVs. (A) The miR-133b expression in neuronal cells following co-culture with BMSCs and treatment of GW4869 or DMSO determined by RT-qPCR. (B) The miR-133b expression in neuronal cells following co-culture with EVs derived from the BMSCs treated with miR-133b inhibitor determined by RT-qPCR. (C) The neuronal cell apoptosis detected by TUNEL assay (× 200). (D) The neuronal cell proliferation detected by CCK-8 assay. (E) The number of NeuN-positive cells in hippocampal CA1 region (× 400). (F) The number of NeuN-positive cells in cerebral cortex (× 400). (G) Apoptosis of neuronal cells in hippocampal CA1 region assessed by TUNEL staining (× 400). (H) Apoptosis of neuronal cells in cerebral cortex assessed by TUNEL staining (× 400). * p vs. the EVs-NC inhibitor group (neuronal cells co-cultured with EVs derived from the BMSCs treated with NC inhibitor). Data were expressed as mean ± standard deviation. Data between two groups were analyzed by unpaired t test while data among multiple groups were analyzed by one-way ANOVA with Tukey's post hoc test. n = 10 in animal experiments. The cell experiments were conducted 3 times independently.

Figure 4. Neuronal cell survival in rats with CA is promoted by miR-133b transferred via BMSCs-derived EVs. (A) The miR-133b expression in neuronal cells following co-culture with BMSCs and treatment of GW4869 or DMSO determined by RT-qPCR. (B) The miR-133b expression in neuronal cells following co-culture with EVs derived from the BMSCs treated with miR-133b inhibitor determined by RT-qPCR. (C) The neuronal cell apoptosis detected by TUNEL assay (× 200). (D) The neuronal cell proliferation detected by CCK-8 assay. (E) The number of NeuN-positive cells in hippocampal CA1 region (× 400). (F) The number of NeuN-positive cells in cerebral cortex (× 400). (G) Apoptosis of neuronal cells in hippocampal CA1 region assessed by TUNEL staining (× 400). (H) Apoptosis of neuronal cells in cerebral cortex assessed by TUNEL staining (× 400). * p < 0.05 vs. the EVs-NC inhibitor group (neuronal cells co-cultured with EVs derived from the BMSCs treated with NC inhibitor). Data were expressed as mean ± standard deviation. Data between two groups were analyzed by unpaired t test while data among multiple groups were analyzed by one-way ANOVA with Tukey's post hoc test. n = 10 in animal experiments. The cell experiments were conducted 3 times independently.