Response Of Neural Stem Cells Research Articles

Abstract WMP42: Remote Ischemic Stroke Decreases Quiescence and Increases Neurogenic Activation of Radial Glia-like Precursors in the Subgranular Zone of the Dentate Gyrus in Adult Mice

Adult neurogenesis occurs in two discrete neurogenic niches in the mammalian brain: the subventricular zone (SVZ) of the lateral ventricles and the subgranular zone (SGZ) of the dentate gyrus in the hippocampal formation. Under physiologic conditions, neurogenesis in these regions is thought to be important for the maintenance and reorganization of interneurons in the olfactory bulb (SVZ), and the modulation and refinement of existing circuits in the dentate gyrus (SGZ). In rodents, stroke enhances neurogenesis in these regions, and stroke-induced neurogenesis appears to play a role in functional recovery. However, little is known about the behavior of quiescent vs. activated pools of neural stem cells in response to stroke, or the mechanisms of stroke-induced neurogenesis at the cellular level. Here we tested the hypothesis that stroke-induced neurogenesis involves the activation of quiescent stem cell pools, either alone or in combination with further differentiation of activated precursors. We used a genetic labeling strategy for in vivo lineage tracing of quiescent, nestin-expressing radial glia-like (RGL) precursors in the SGZ to elucidate the effect of remote focal ischemic infarction on RGLs at a clonal level. Unilateral infarction of the visual cortex by photothrombosis in transgenic mice caused an increase in the number of differentiated neuronal precursors and a decrease in the number of quiescent RGLs in the ipsilateral SGZ. Furthermore, among activated RGL clones, there was an increase in symmetric and neurogenic RGL divisions in response to stroke, with a concomitant loss of astrogliogenic RGL divisions. These preliminary results suggest that stroke may alter hippocampal neurogenesis from a distance by activating quiescent RGL pools in the SGZ through an as-of-yet unidentified mechanism and pushing them toward a neuronal fate. In turn, this may accelerate the time-dependent depletion of this RGL population. We speculate that this phenomenon contributes to cognitive changes associated with stroke and may represent a target for pharmacological intervention.

DNA methyltransferase mediates dose-dependent stimulation of neural stem cell proliferation by folate

The proliferative response of neural stem cells (NSCs) to folate may play a critical role in the development, function and repair of the central nervous system. It is important to determine the dose-dependent effects of folate in NSC cultures that are potential sources of transplantable cells for therapies for neurodegenerative diseases. To determine the optimal concentration and mechanism of action of folate for stimulation of NSC proliferation in vitro, NSCs were exposed to folic acid or 5-methyltetrahydrofolate (5-MTHF) (0–200 μmol/L) for 24, 48 or 72 h. Immunocytochemistry and methyl thiazolyl tetrazolium assay showed that the optimal concentration of folic acid for NSC proliferation was 20–40 μmol/L. Stimulation of NSC proliferation by folic acid was associated with DNA methyltransferase (DNMT) activation and was attenuated by the DNMT inhibitor zebularine, which implies that folate dose-dependently stimulates NSC proliferation through a DNMT-dependent mechanism. Based on these new findings and previously published evidence, we have identified a mechanism by which folate stimulates NSC growth.

Patterning of human cord blood-derived stem cells on single cell posts and lines: Implications for neural commitment.

Using stem cells (SC) in new strategies for clinical treatment requires control of stem cell fate decision and the ability to govern their patterning and commitment in tissue engineering. Neural stem cells and other adult SC can respond to the different components of the microenvironment and their spatial arrangement in the stem cell niche. It has been shown previously by our group that different composition and architecture of patterned bioactive domains influence the developmental response of neural stem cells. In the present report we verify the commitment and differentiation of neural stem cells derived from human cord blood (HUCB-NSC) by a single cell patterning system. Microcontact printing technology was used to generate single cell positioning areas of different geometry: 10 micrometer-thin lines and 10 micrometer-width one cell posts. The commitment and differentiation of HUCB-NSC cells cultured on different surfaces were dependent on the geometry and the type of biomaterial present in bioactive domains. Fibronectin promoted neuronal protrusion outgrowth (Beta-tubulin III and MAP-2 positive cells) and gap junction development (Cx43 marker) between cells growing on lines while poly-L-lysine promoted HUCB-NSC differentiation into astrocytic, glial fibrillary acidic protein expressing (GFAP positive) cell phenotype. Here we also demonstrate by scanning electron microscopy that morphology of cells on the patterned surface is highly dependent upon the type of biomolecules used for printing. These kinds of platforms can be used for investigating the influence of spatial organization of environment on the SC fate decision and for studying the molecular processes occurring in a single cell.

Changes in the Neurogenesis and Axonal Sprouting in the Organotypic Hippocampal Slice Culture by Aβ25-35Treatment

Induction of neurogenesis can occur in the hippocampus in response to various pathological conditions, such as Alzheimer's disease. The aim of this study was to investigate the changes that occur in endogenous neural stem cells in response to amyloid beta -induced neuronal cell damage in organotypic hippocampal slice cultures. Cresyl violet staining and Fluoro-Jade B staining were used to detect neuronal cell damage and changes of mossy fiber terminals were observed by Timm's staining. The immunofl uorescence staining was used to detect the newly generated cells in the subgranular zone (SGZ) of the dentate gyrus with specific marker, 5-bromo-2'-deoxyuridine (BrdU), Ki-67, Nestin, and doublecortin (DCX). In compared to control slices, neuronal cell damage was observed and the mossy fibers were expanded to CA3 area by treatment with . Ki-67/Nestin- and BrdU/DCX-positive cells were detected in the SGZ. In conclusion, these results demonstrate that -induced neuronal damage results in an increase in endogenous neural stem cells in rat hippocampal slice cultures not only for gliosis but also for neurogenesis.

Neuronal circuitry mechanism regulating adult quiescent neural stem-cell fate decision

Adult neurogenesis arises from neural stem cells within specialized niches1–3. Neuronal activity and experience, presumably acting upon this local niche, regulate multiple stages of adult neurogenesis, from neural progenitor proliferation to new neuron maturation, synaptic integration and survival1, 3. Whether local neuronal circuitry has a direct impact on adult neural stem cells is unknown. Here we show that in the adult hippocampus nestin-expressing radial glia-like quiescent neural stem cells4–9 (RGLs) respond tonically to the neurotransmitter GABA via γ2 subunit-containing GABAA Rs. Clonal analysis9 of individual RGLs revealed a rapid exit from quiescence and enhanced symmetric self-renewal after conditional γ2 deletion. RGLs are in close proximity to GAD67+ terminals of parvalbumin-expressing (PV+) interneurons and respond tonically to GABA released from these neurons. Functionally, optogenetic control of dentate PV+, but not somatostatin- or vasoactive intestinal polypeptide (VIP)-expressing, interneuron activity can dictate the RGL choice between quiescence and activation. Furthermore, PV+ interneuron activation restores RGL quiescence following social isolation, an experience that induces RGL activation and symmetric division8. Our study identifies a niche cell-signal-receptor trio and a local circuitry mechanism that control the activation and self-renewal mode of quiescent adult neural stem cells in response to neuronal activity and experience.

Highlights from Recent Cancer Literature

Highlights from Recent Cancer Literature

Distinct Neural Stem Cell Populations Give Rise to Disparate Brain Tumors in Response to N-MYC

The proto-oncogene MYCN is mis-expressed in various types of human brain tumors. To clarify how developmental and regional differences influence transformation, we transduced wild-type or mutationally stabilized murine N-myc(T58A) into neural stem cells (NSCs) from perinatal murine cerebellum, brain stem, and forebrain. Transplantation of N-myc(WT) NSCs was insufficient for tumor formation. N-myc(T58A) cerebellar and brain stem NSCs generated medulloblastoma/primitive neuroectodermal tumors, whereas forebrain NSCs developed diffuse glioma. Expression analyses distinguished tumors generated from these different regions, with tumors from embryonic versus postnatal cerebellar NSCs demonstrating Sonic Hedgehog (SHH) dependence and SHH independence, respectively. These differences were regulated in part by the transcription factor SOX9, activated in the SHH subclass of human medulloblastoma. Our results demonstrate context-dependent transformation of NSCs in response to a common oncogenic signal.

Abstract 3588: Induced Pluripotent and Fetal Derived Neural Stem Cells Differ in Chemokine and Receptor Expression.

Introduction: Cell based therapies have shown a considerable ability to improve functional outcome when administered after experimental stroke. Intra-arterial (IA) cell transplantation has been shown to be a valuable alternative transplantation method for cell treatment. The success of IA therapy depends on the targeted homing and subsequent migration of cells to the injured brain area. This process is reliant on neural stem cell (NSC) expression of chemokines and their receptors. Understanding the differences in cell expression of these molecules and the resulting differences in chemotaxis will help identify ideal cell types for transplantation. Previously, we compared mRNA from human and mouse NSCs and found that only a 10% overlap in chemokine expression. We wanted to determine if there are also variations between different cell types from the same species. We compared NSCs obtained from induced pluripotent stem cells (iPS NSCs), to fetal NSCs (F NSC). We used RT-qPCR, in vitro cell migration, and quantification of cell secretion to compare chemokine and ligand expression in F NSCs and iPS NSCs. Methods: Secondary neurospheres originating from FVB mice were generated from either fetal primary NSCs from E17.5 mice (F NSC), or derived by transducing neurospheres originating from E3 mice with Oct4 (iPS NSC). Cells were grown as neurospheres in growth media treated with EGF and FGF. RT-qPCR was performed on extracted RNA using microarray technology for chemokine receptors and ligands. Luminex immunological assay was completed to assess secreted cytokines and inflammatory proteins. The Boyden chamber migration assay was used to determine migration to SDF-1 or MCP-1 at varying concentrations. Results: RNA expression was detected for 58% of the factors assessed. F NSCs alone expressed 6% (e.g. mmp2, Cxcl10), iPS NSCs alone expressed 16% (e.g. CCL2, CCL7, LIF), and both iPS NSCs and F NSCs expressed 20% (e.g. BDNF, HIF1a, CXCR4) of the detected factors. We assessed 26 secreted proteins, 73% occurred at detectable levels, of these 74% were expressed more highly in F NSCs than in iPS NSCs, including, il-1α, CCL3, CCL5, TNF-α, and VEGF; iPS NSCs secreted higher levels of MCP3, Il-3, and substantially more MCP-1 (113.7 ± 1.44 vs 52.78 ±4.221 pg/ml). iPS NSCs also had a higher migratory response to MCP-1 than F NSCs did (p < 0.001). Cell migration to SDF-1 did not differ between cell types (p = 0.905). Conclusions: Several chemokines and their receptors were differentially expressed in mouse iPS and F NSCs. These differences may result in variations in the NSC response to the ischemic brain. The profile of chemokine and chemokine receptor expression on NSCs should be strongly considered when selecting cell types for post stroke treatment.

Critical Role of PI3K/Akt/GSK3β in Motoneuron Specification from Human Neural Stem Cells in Response to FGF2 and EGF

Fibroblast growth factor (FGF) and epidermal growth factor (EGF) are critical for the development of the nervous system. We previously discovered that FGF2 and EGF had opposite effects on motor neuron differentiation from human fetal neural stem cells (hNSCs), but the underlying mechanisms remain unclear. Here, we show that FGF2 and EGF differentially affect the temporal patterns of Akt and glycogen synthase kinase 3 beta (GSK3β) activation. High levels of phosphatidylinositol 3-kinase (PI3K)/Akt activation accompanied with GSK3β inactivation result in reduction of the motor neuron transcription factor HB9. Inhibition of PI3K/Akt by chemical inhibitors or RNA interference or overexpression of a constitutively active form of GSK3β enhances HB9 expression. Consequently, PI3K inhibition increases hNSCs differentiation into HB9+/microtubule-associated protein 2 (MAP2)+ motor neurons in vitro. More importantly, blocking PI3K not only enhances motor neuron differentiation from hNSCs grafted into the ventral horn of adult rat spinal cords, but also permits ectopic generation of motor neurons in the dorsal horn by overriding environmental influences. Our data suggest that FGF2 and EGF affect the motor neuron fate decision in hNSCs differently through a fine tuning of the PI3K/AKT/GSK3β pathway, and that manipulation of this pathway can enhance motor neuron generation.

The CCR2/CCL2 Interaction Mediates the Transendothelial Recruitment of Intravascularly Delivered Neural Stem Cells to the Ischemic Brain

The inflammatory response is a critical component of ischemic stroke. In addition to its physiological role, the mechanisms behind transendothelial recruitment of immune cells also offer a unique therapeutic opportunity for translational stem cell therapies. Recent reports have demonstrated homing of neural stem cells (NSC) into the injured brain areas after intravascular delivery. However, the mechanisms underlying the process of transendothelial recruitment remain largely unknown. Here we describe the critical role of the chemokine CCL2 and its receptor CCR2 in targeted homing of NSC after ischemia. Twenty-four hours after induction of stroke using the hypoxia-ischemia model in mice CCR2+/+ and CCR2-/- reporter NSC were intra-arterially delivered. Histology and bioluminescence imaging were used to investigate NSC homing to the ischemic brain. Functional outcome was assessed with the horizontal ladder test. Using NSC isolated from CCR2+/+ and CCR2-/- mice, we show that receptor deficiency significantly impaired transendothelial diapedesis specifically in response to CCL2. Accordingly, wild-type NSC injected into CCL2-/- mice exhibited significantly decreased homing. Bioluminescence imaging showed robust recruitment of CCR2+/+ cells within 6 hours after transplantation in contrast to CCR2-/- cells. Mice receiving CCR2+/+ grafts after ischemic injury showed a significantly improved recovery of neurological deficits as compared to animals with transplantation of CCR2-/- NSC. The CCL2/CCR2 interaction is critical for transendothelial recruitment of intravascularly delivered NSC in response to ischemic injury. This finding could have significant implications in advancing minimally invasive intravascular therapeutics for regenerative medicine or cell-based drug delivery systems for central nervous system diseases.

Deafferentation enhances neurogenesis in the young and middle aged hippocampus but not in the aged hippocampus

Increased neurogenesis in the dentate gyrus (DG) after brain insults such as excitotoxic lesions, seizures, or stroke is a well known phenomenon in the young hippocampus. This plasticity reflects an innate compensatory response of neural stem cells (NSCs) in the young hippocampus to preserve function or minimize damage after injury. However, injuries to the middle-aged and aged hippocampi elicit either no or dampened neurogenesis response, which could be due to an altered plasticity of NSCs and/or the hippocampus with age. We examined whether the plasticity of NSCs to increase neurogenesis in response to a milder injury such as partial deafferentation is preserved during aging. We quantified DG neurogenesis in the hippocampus of young, middle-aged, and aged F344 rats after partial deafferentation. A partial deafferentation of the left hippocampus without any apparent cell loss was induced via administration of Kainic acid (0.5 μg in 1.0 μl) into the right lateral ventricle of the brain. In this model, degeneration of CA3 pyramidal neurons and dentate hilar neurons in the right hippocampus results in loss of commissural axons which leads to partial deafferentation of the dendrites of dentate granule cells and CA1-CA3 pyramidal neurons in the left hippocampus. Quantification of newly born cells that are added to the dentate granule cell layer at postdeafferentation days 4-15 using 5'-bromodeoxyuridine (BrdU) labeling revealed greatly increased addition of newly born cells (∼three fold increase) in the deafferented young and middle-aged hippocampi but not in the deafferented aged hippocampus. Measurement of newly born neurons using doublecortin (DCX) immunostaining also revealed similar findings. Analyses using BrdU-DCX dual immunofluorescence demonstrated no changes in neuronal fate-choice decision of newly born cells after deafferentation, in comparison to the age-matched naive hippocampus in all age groups. Thus, the plasticity of hippocampal NSCs to increase DG neurogenesis in response to a milder injury such as partial hippocampal deafferentation is preserved until middle age but lost at old age.

Stage-dependent STAT3 activation is involved in the differentiation of rat hippocampus neural stem cells

Extracts of deafferented hippocampus were previously found to promote neuronal differentiation of neural stem cells (NSCs). To gain insights into the underlying molecular mechanisms we studied the potential involvement of signal transducer and activator of transcription3 (STAT3) activation in the NSCs response to hippocampal extracts. Here we report that phosphorylated STAT3 (p-STAT3) is expressed at different stages in neurons and astrocytes differentiated from rat hippocampus-derived NSCs. Deafferented hippocampal extracts produced sustained upregulation of p-STAT3 levels and promoted NSC differentiation and neurogenesis, whereas extracts of normal hippocampus were without effect. Interleukin-6 (IL-6), an activator of JAK/STAT signaling pathways, had no effect on neurogenesis, whereas the selective STAT3 inhibitor p-ip-STAT3 decreased the number of Microtubule-associated protein-2 (MAP-2)-positive cells generated by NSC differentiation. These findings argue that STAT3-related signaling pathways are likely to play a role in neuronal survival and differentiation during NSC neurogenesis stimulated by extracts of deafferented hippocampus.

Severe Hypoxia: Consequences to Neural Stem Cells and Neurons

Multiple neurological diseases result from a pathological hypoxia in the brain, resulting in various motor, sensory or cognitive sequelae. Understanding the response of neural stem cells (NSCs) and differentiated neurons to hypoxia will help better treat such diseases. We exposed mouse embryonic primary neurons (PN) and neural stem cells to 1% O2 in vitro. Both cell types survived and retained their immunocyto-chemical markers, and neurons showed no obvious morphological changes. Microarray analysis showed that the number of genes with significantly altered expression levels was almost five-fold higher in NSCs compared to PN. NSCs displayed a clear block in G1/S phase of the cell cycle and a number of down-regulated cytokine genes. Various growth factors (e.g. neural growth factor, prolactin), involved in survival and proliferation, genes of the Notch pathway, and genes involved in glial differentiation, and cell-matrix adhesion were up-regulated. PN displayed a down-regulation of a number of genes involved in neuron-specific functions, in particular, transmitter-related (e.g. synaptic transmission, neurotransmitter transport and release, learning, adult behavior). We conclude that hypoxia 1-down-regulates genes involved in multiple neuronal functions which can negatively impact learning and memory; 2-induces a cell cycle block in NSCs; 3-can precondition NSC towards a particular differentiation potential while maintaining them fully undifferentiated.

Lysosome Dysfunction Triggers Atg7-dependent Neural Apoptosis

Macroautophagy (autophagy) is a process wherein bulk cytosolic proteins and damaged organelles are sequestered and degraded via the lysosome. Alterations in autophagy-associated proteins have been shown to cause neural tube closure defects, neurodegeneration, and tumor formation. Normal lysosome function is critical for autophagy completion and when altered may lead to an accumulation of autophagic vacuoles (AVs) and caspase activation. The tumor suppressor p53 is highly expressed in neural precursor cells (NPCs) and has an important role in the regulation of both autophagy and apoptosis. We hypothesized that altered lysosome function would lead to NPC death via an interaction between autophagy- and apoptosis-associated proteins. To test our hypothesis, we utilized FGF2-expanded NPCs and the neural stem cell line, C17.2, in combination with the lysosomotropic agent chloroquine (CQ) and the vacuolar ATPase inhibitor bafilomycin A1 (Baf A1). Both CQ and Baf A1 caused concentration- and time-dependent AV accumulation, p53 phosphorylation, increased damage regulator autophagy modulator levels, caspase-3 activation, and cell death. Short hairpin RNA knockdown of Atg7, but not Beclin1, expression significantly inhibited CQ- and Baf A1-induced cell death, indicating that Atg7 is an upstream mediator of lysosome dysfunction-induced cell death. Cell death and/or caspase-3 activation was also attenuated by protein synthesis inhibition, p53 deficiency, or Bax deficiency, indicating involvement of the intrinsic apoptotic death pathway. In contrast to lysosome dysfunction, starvation-induced AV accumulation was inhibited by either Atg7 or Beclin1 knockdown, and Atg7 knockdown had no effect on starvation-induced death. These findings indicate that Atg7- and Beclin1-induced autophagy plays a cytoprotective role during starvation but that Atg7 has a unique pro-apoptotic function in response to lysosome dysfunction.

A protein interaction network for the analysis of the neuronal differentiation of neural stem cells in response to titanium dioxide nanoparticles

The effects of titanium dioxide (TiO 2) nanoparticles (NPs) on the differentiation of neural stem cells are reported. Our findings indicate that TiO 2 NPs lead to a differentiational tendency towards neurons from neural stem cells, suggesting TiO 2 NPs might be a beneficial inducer for neuronal differentiation. To insight into the possible molecular mechanism of the neuronal differentiation, we conducted a protein–protein interaction network (PIN) analysis. To this end, a global mapping of target proteins induced by TiO 2 NPs was first made by a 2-dimensional electrophoresis analysis. Results showed that 9 proteins were significantly changed and then they were subjected to the mass spectrometric assay. All 9 identified proteins are involved in signal, molecular chaperones, cytoskeleton, and nucleoprotein. Further, based on our experimental data and DIP, IntAct–EBI, GRID database, a protein–protein interaction network was constructed, which provides highly integrated information exhibiting the protein–protein interaction. By analysis of the gene expression, the signal pathway involving Cx43 phosphorylation, which is negatively regulated by the protein kinase C epsilon (PKCɛ), is demonstrated. It is inferred that PKCɛ plays a pivotal negative role in the neuronal differentiation of stem neural cells in response to the TiO 2 NPs exposure.

Neurotoxicity Screening in a Multipotent Neural Stem Cell Line Established from the Mouse Brain

Neural stem cells (NSCs) have mainly been applied to neurodegeneration in some medically intractable neurologic diseases. In this study, we established a novel NSC line and investigated the cytotoxic responses of NSCs to exogenous neurotoxicants, glutamates and reactive oxygen species (ROS). A multipotent NSC line, B2A1 cells, was established from long-term primary cultures of oligodendrocyte-enriched cells from an adult BALB/c mouse brain. B2A1 cells could be differentiated into neuronal, astrocytic and oligodendroglial lineages. The cells also expressed genotypic mRNA messages for both neural progenitor cells and differentiated neuronoglial cells. B2A1 cells treated with hydrogen peroxide and L-buthionine-(S,R)-sulfoximine underwent 30-40% cell death, while B2A1 cells treated with glutamate and kainate showed 25-35% cell death. Cytopathologic changes consisting of swollen cell bodies, loss of cytoplasmic processes, and nuclear chromatin disintegration, developed after exposure to both ROS and excitotoxic chemicals. These results suggest that B2A1 cells may be useful in the study of NSC biology and may constitute an effective neurotoxicity screening system for ROS and excitotoxic chemicals.

AMP-activated Protein Kinase Is Involved in Neural Stem Cell Growth Suppression and Cell Cycle Arrest by 5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside and Glucose Deprivation by Down-regulating Phospho-retinoblastoma Protein and Cyclin D

The fate of neural stem cells (NSCs), including their proliferation, differentiation, survival, and death, is regulated by multiple intrinsic signals and the extrinsic environment. We had previously reported that 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) directly induces astroglial differentiation of NSCs by activation of the Janus kinase (JAK)/Signal transducer and activator of transcription 3 (STAT3) pathway independently of AMP-activated protein kinase (AMPK). Here, we reported the observation that AICAR inhibited NSC proliferation and its underlying mechanism. Analysis of caspase activity and cell cycle showed that AICAR induced G1/G0 cell cycle arrest in NSCs, associated with decreased levels of poly(ADP-ribose) polymerase, phospho-retinoblastoma protein (Rb), and cyclin D but did not cause apoptosis. Iodotubericidin and Compound C, inhibitors of adenosine kinase and AMPK, respectively, or overexpression of a dominant-negative mutant of AMPK, but not JAK inhibitor, were able to reverse the anti-proliferative effect of AICAR. Glucose deprivation also activated the AMPK pathway, induced G0/G1 arrest, and suppressed the proliferation of NSCs, an effect associated with decreased levels of phospho-Rb and cyclin D protein. Furthermore, Compound C and overexpression of dominant-negative AMPK in C17.2 NSCs could block the glucose deprivation-mediated down-regulation of cyclin D and partially reverse the suppression of proliferation. These results suggest that AICAR and glucose deprivation might induce G1/G0 cell cycle arrest and suppress proliferation of NSCs via phospho-Rb and cyclin D down-regulation. AMPK, but not JAK/STAT3, activation is key for this inhibitory effect and may play an important role in the responses of NSCs to metabolic stresses such as glucose deprivation.

In vitro behavior of neural stem cells in response to different chemical functional groups

Neural stem cells (NSCs) cultured on glass surfaces modified by different chemical groups, including hydroxyl (–OH), sulfonic (–SO 3H), amino (–NH 2), carboxyl (–COOH), mercapto (–SH) and methyl (–CH 3) groups, are shown here to commit to phonotypes with extreme sensitivity to surface chemical groups. The adhering NSCs at the level of single cells exhibited morphological changes in response to different chemical groups. NSCs on –SO 3H surfaces had the largest contact area and the most flattened morphology, while those on –CH 3 surfaces exhibited the smallest contact area and the most rounded morphology. After 5 days of culture, the migration of NSCs from their original aggregates onto these test surfaces followed the trend: –NH 2>–COOH = –SH≫–SO 3H>–CH 3>–OH. The expression of specific markers, including nestin, β-Tubulin-III, glial fibrillary acidic protein and O4, were used to examine NSCs lineage specification. The –SO 3H surfaces favored NSCs differentiation into oligodendrocytes, while NSCs in contact with –COOH, –NH 2, –SH and –CH 3 had the ability to differentiate into neurons, astrocytes and oligodendrocytes. Compared to –COOH surfaces, –NH 2 seemed to promote neuronal differentiation. These chemically modified surfaces exhibited regulation of NSCs on adhesion, migration and differentiation potential, providing chemical means for the design of biomaterials to direct NSCs lineage specification for neural tissue engineering.

Gold ions bio-released from metallic gold particles reduce inflammation and apoptosis and increase the regenerative responses in focal brain injury

Traumatic brain injury results in loss of neurons caused as much by the resulting neuroinflammation as by the injury. Gold salts are known to be immunosuppressive, but their use are limited by nephrotoxicity. However, as we have proven that implants of pure metallic gold release gold ions which do not spread in the body, but are taken up by cells near the implant, we hypothesize that metallic gold could reduce local neuroinflammation in a safe way. Bio-liberation, or dissolucytosis, of gold ions from metallic gold surfaces requires the presence of disolycytes i.e. macrophages and the process is limited by their number and activity. We injected 20-45 mum gold particles into the neocortex of mice before generating a cryo-injury. Comparing gold-treated and untreated cryolesions, the release of gold reduced microgliosis and neuronal apoptosis accompanied by a transient astrogliosis and an increased neural stem cell response. We conclude that bio-liberated gold ions possess pronounced anti-inflammatory and neuron-protective capacities in the brain and suggest that metallic gold has clinical potentials. Intra-cerebral application of metallic gold as a pharmaceutical source of gold ions represents a completely new medical concept that bypasses the blood-brain-barrier and allows direct drug delivery to inflamed brain tissue.

Glial fibrillary acidic protein‐expressing cells in the neurogenic regions in normal and injured adult brains

In the adult brain, neurogenic stem cells are prevalent in the subventricular zone (SVZ) of the lateral ventricle wall and the subgranular zone (SGZ) in the dentate gyrus. Cells that have structural and molecular characteristics of astrocytes function as neurogenic stem cells in these regions, in which these cells also participate in the creation of the microenvironment that stimulates neurogenesis. In the present paper, we review the phenotypic properties, subpopulations, and proliferation of glial fibrillary acidic protein (GFAP)-expressing cells in these two neurogenic regions and their responses to different brain injuries. Cells fulfilling the criteria for astrocytes, i.e., expressing GFAP, in the SVZ and SGZ respond differently to brain injuries or neurogenic stimuli. The importance of guidance by astrocytes of newly formed neuronal cells is emphasized. The assessment of GFAP-expressing cells in the neurogenic regions is of great importance for understanding the mechanism underlying the response of neural stem cells to brain injury.