Journal: Neural Regeneration Research

Article Title: Exosomes originating from neural stem cells undergoing necroptosis participate in cellular communication by inducing TSC2 upregulation of recipient cells following spinal cord injury

doi: 10.4103/NRR.NRR-D-24-00068

Figure Lengend Snippet: Validation of the necroptosis model and characterization of NSCs and exosomes. (A) Characterization of NSC neurospheres. The NSCs displayed a typical spherical neurosphere morphology. Scale bars: 500 μm (upper) and 100 μm (lower). (B) Representative immunofluorescence images of SOX2 (green, Alexa Fluor 488) and Nestin (red, Alexa Fluor 594) expression in NSCs. Both neurospheres and individual NSCs expressed SOX2 and Nestin. Cell nuclei were counterstained with DAPI (blue). Scale bar: 100 μm. (C) NSCs were treated with DMSO or TSZ for 8 hours, after which cell death was analyzed by flow cytometry. (D) Exosome morphology as assessed by transmission electron microscopy. The exosomes in both the NSC-control and NSC-TSZ groups exhibited a typical one-sided semi-concave bilayer membrane structure. Scale bars: 200 nm. (E, F) Nanoflow cytometry detection of particle size distribution, concentration, and exosome surface marker expression. DAPI: 4′,6-Diamidino-2-phenylindole; DMSO: dimethyl sulfoxide; NSC: neural stem cell; SOX2: SRY-box transcription factor 2; TSZ: tumor necrosis factor α, SMAC mimetic, zVAD-fmk.

Article Snippet: NSCs were pre-treated with 20 μM zVAD-fmk (MCE, Monmouth Junction, NJ, USA) for 30 minutes, then treated with recombinant mouse tumor necrosis factor α (TNFα, 40 ng/mL, Peprotech) and 4 μM SMAC mimetic (MCE) for 6 hours to establish a TSZ (TNFα, SMAC mimetic, zVAD-fmk)-induced in vitro model of necroptosis.

Techniques: Immunofluorescence, Expressing, Flow Cytometry, Transmission Assay, Electron Microscopy, Control, Membrane, Cytometry, Concentration Assay, Marker

Journal: Neural Regeneration Research

Article Title: Exosomes originating from neural stem cells undergoing necroptosis participate in cellular communication by inducing TSC2 upregulation of recipient cells following spinal cord injury

doi: 10.4103/NRR.NRR-D-24-00068

Figure Lengend Snippet: Western blot and immunofluorescence analyses of TSC2 expression after uptake of exosomes derived from TSZ-treated NSCs in vitro . (A, B) Western blot and semi-quantitative analyses of TSC2 expression at different time points after uptake of exosomes derived from TSZ-treated NSCs in vitro . Data are expressed as mean ± SD ( n = 3), and were analyzed by one-way analysis of variance followed by Bonferroni’s post hoc test. (C) Immunofluorescence staining for TSC2 (green, Alexa Fluor 488) and DID-labeled exosomes derived from TSZ-treated NSCs (red) 12 hours after uptake in vitro . When the NSC-34 cells took up the exosomes, there was a significant increase in TSC2 expression. Cell nuclei were counterstained with DAPI. Scale bars: 50 μm (left) and 20 μm (middle and right). DAPI: 4′,6-Diamidino-2-phenylindole; DID: 1,1-dioctadecyl-3,3,3,3-tetramethylindodicarbocyanine; TSC2: tuberous sclerosis 2; TSZ: tumor necrosis factor α, SMAC mimetic, zVAD-fmk.

Article Snippet: NSCs were pre-treated with 20 μM zVAD-fmk (MCE, Monmouth Junction, NJ, USA) for 30 minutes, then treated with recombinant mouse tumor necrosis factor α (TNFα, 40 ng/mL, Peprotech) and 4 μM SMAC mimetic (MCE) for 6 hours to establish a TSZ (TNFα, SMAC mimetic, zVAD-fmk)-induced in vitro model of necroptosis.

Techniques: Western Blot, Immunofluorescence, Expressing, Derivative Assay, In Vitro, Staining, Labeling

Journal: Neural Regeneration Research

Article Title: Enhanced neurogenesis after ischemic stroke: The interplay between endogenous and exogenous stem cells

doi: 10.4103/NRR.NRR-D-24-00879

Figure Lengend Snippet: The endogenous heterogeneity of NSCs and stroke recovery. Diagram showing two major NSC niches SVZ/SGZ and the features of different subpopulations of NSCs. Under physiological conditions, NSCs in these two regions continuously generate new cells to maintain normal functions in the olfactory bulb and hippocampus. After an ischemic stroke, NSCs from these areas also contribute to neurorestoration (Lim and Alvarez-Buylla, 2016; Abbott and Nigussie, 2020). The materials were sourced from Servier Medical Art ( http://smart.servier.com/ ), licensed under a Creative Commons Attribution 3.0 Generic License ( https://creativecommons.org/licenses/by/3.0/ ). Ascl1: Achaete-scute complex-Like 1; BDNF: brain-derived neurotrophic factor; BMPs: bone morphogenetic proteins; DCX: doublecortin; GABARs: gamma-aminobutyric acid receptors; GFAP: glial fibrillary acidic protein; IGF-1: insulin-like growth factor-1; NSC: neural stem cell; RMS: rostral migration stream; SGZ: subgranular zone; Sox2: sex-determining region Y-box-2; VCAM1: vascular cell adhesion protein 1.

Article Snippet: Besides forming natural scaffolds (biobridges) and secreting neuron-supporting cytokines, chemokines, and growth factors, exogenous stem cells may also stimulate the survival, proliferation, migration, and differentiation of endogenous NSCs by modifying the environment in the ischemic area.

Techniques: Derivative Assay, Migration

Journal: Neural Regeneration Research

Article Title: Enhanced neurogenesis after ischemic stroke: The interplay between endogenous and exogenous stem cells

doi: 10.4103/NRR.NRR-D-24-00879

Figure Lengend Snippet: Modulation of exogenous stem cells on endogenous NSCs. The schematic illustrates the three primary actions of exogenous stem cells on endogenous NSCs. Exogenous stem cells can promote the proliferation of endogenous NSCs through cytokines (such as BDNF, EGF, and IGF), exosomes, and direct contact. They can also recruit endogenous NSCs to migrate to the damaged area by establishing a “biobridge.” Additionally, exogenous stem cells can indirectly improve the microenvironment following ischemia, enhancing the survival and differentiation of endogenous NSCs. By targeting common pathways in both endogenous NSCs and exogenous stem cells, certain drugs, hydrogels, and exosomes can synergize with stem cell transplantation to improve neurological function recovery more effectively. The materials were sourced from Servier Medical Art ( http://smart.servier.com/ ), licensed under a Creative Commons Attribution 3.0 Generic License. ( https://creativecommons.org/licenses/by/3.0/ ). BDNF: Brain-derived neurotrophic factor; EGF: epidermal growth factor; ES: embryonic stem cells; EVs: extracellular vesicles; IGF: insulin-like growth factor; IL: interleukin; iPSC: induced pluripotent stem cells; MMP-9: matrix metalloproteinase 9; MSCs: mesenchymal stem cells; NGF: nerve growth factor; NPCs: neural progenitor cells; NSCs: neural stem cells; PDGF: platelet-derived growth factor; TNF-α: tumor necrosis factor-alpha; TNF-β: tumor necrosis factor-beta; VEGF: vascular endothelial growth factor.

Article Snippet: Besides forming natural scaffolds (biobridges) and secreting neuron-supporting cytokines, chemokines, and growth factors, exogenous stem cells may also stimulate the survival, proliferation, migration, and differentiation of endogenous NSCs by modifying the environment in the ischemic area.

Techniques: Transplantation Assay, Derivative Assay

Journal: Neural Regeneration Research

Article Title: Targeting capabilities of engineered extracellular vesicles for the treatment of neurological diseases

doi: 10.4103/NRR.NRR-D-24-00462

Figure Lengend Snippet: Clinical trials investigating EVs for neurological diseases

Article Snippet: ArunA Biomedical , NSC-EV , IS , Preclinical/Webb et al., 2018.

Techniques:

Journal: Neural Regeneration Research

Article Title: Human induced pluripotent stem cell–derived therapies for regeneration after central nervous system injury

doi: 10.4103/NRR.NRR-D-24-00901

Figure Lengend Snippet: Summary of transplant studies

Article Snippet: Wu et al., 2021: Nerve growth factor (NGF) with hypoxia response elements loaded by adeno-associated virus (AAV) combined with neural stem cells improve the spinal cord injury recovery , Isolated Sprague- Dawley NSC 5HRE-NGF , Sprague-Dawley adult female (220-250 g) , T9 impact SCI (10 g hammer, 25 mm height) , PBS injection , 7 d post-SCI , 2.0×10 5 NSCs cells/10 μL , Improved locomotion and hindlimb function. Downregulation of autophagy- induced cell death.

Techniques: Transplantation Assay, Animal Model, Cell Counting, Virus, Isolation, Injection, Derivative Assay, Microinjection, Functional Assay, Migration, Expressing, Knock-In, Activity Assay, Control, Optogenetics, Inhibition, Saline, Shear, Encapsulation, Anterograde Tracing, In Vivo, Patch Clamp, In Vitro, Staining