• Altricial
• Grasscutter
• Greater cane rat
• Neocortex development
• Neurogenesis
• Neuron maturation
• Precocial

• Animals
• Female
• Pregnancy
• Rats
• Neocortex
• Neural Stem Cells/metabolism
• Neurogenesis/physiology
• Neurons/metabolism
• Rodentia

• International Society for Neurochemistry (ISN)-CAEN Category 1A Grants 2017
• Needs Assessment Intervention Fund of the Federal Government of Nigeria
• Universität Leipzig (1039)

• Cell-based therapies
• Clinical trials
• Neural stem cells
• Regenerative medicine
• Spinal cord injury
• Transplantation

• adult human neural stem cells
• middle cerebral artery occlusion
• neuroprotection
• pro-angiogenic effects
• stroke

• Humans
• Brain/cytology
• Cell Culture Techniques/methods
• Neurons/cytology
• Adult
• Cells, Cultured
• Neurosurgical Procedures/methods

• Brain bank
• Human brain
• Neural progenitor cell
• Neurosphere

• Bioengineering
• Cell therapy
• Disability
• Scaffold
• Trauma

• Animals
• Brain/pathology
• Brain Tissue Transplantation
• Cell Survival
• Doublecortin Domain Proteins
• Epilepsy/pathology
• Genetic Vectors/metabolism
• Glial Fibrillary Acidic Protein/metabolism
• Green Fluorescent Proteins/metabolism
• Humans
• Lentivirus/metabolism
• Male
• Microtubule-Associated Proteins/metabolism
• Neural Stem Cells/metabolism
• Neuropeptides/metabolism
• Peptides/chemistry
• Rats, Wistar
• Recovery of Function
• Spinal Cord Injuries/pathology
• Tissue Scaffolds/chemistry
• Transduction, Genetic
• Rats

• 964650 National Institute for Medical Research Development

• altricial
• cortex development
• dwarf rabbit
• guinea pig
• neurogenesis
• neuron maturation
• precocial

• Jurkat cells
• electrophysiology
• human induced pluripotent stem cell-derived neurons
• patch clamping
• temperature control

• Center for Ultrafast Imaging (CUI) under the Excellence Program (EXC-1024) Deutsche Forschungsgemeinschaft
• Priority Program (SPP 1857) ESSENCE Deutsche Forschungsgemeinschaft
• _ Deutscher Akademischer Austauschdienst
• BioPict Joachim Herz Stiftung

Nanostructured substrates such as nanowire arrays form a powerful tool for building next-generation medical devices.

Primary neurons from rodent brain hippocampus and cortex have served as important tools in biomedical research over the years. However, protocols for the preparation of primary neurons vary, which often lead to conflicting results. This report provides a robust and reliable protocol for the production of primary neuronal cultures from the cortex and hippocampus with minimal contribution of non-neuronal cells. The neurons were grown in serum-free media and maintained for several weeks without any additional feeder cells. The neuronal cultures maintained according to this protocol differentiate and by 3 weeks develop extensive axonal and dendritic branching. The cultures produced by this method show excellent reproducibility and can be used for histological, molecular and biochemical methods.

• cerebral cortex
• hippocampus
• primary neuron

• Excitability
• Induced pluripotent stem cells
• Neuronal maturation
• Rho kinase
• Y-27632

• Amides/pharmacology
• Cell Shape/drug effects
• Electrophysiological Phenomena/drug effects
• Humans
• Induced Pluripotent Stem Cells/cytology
• Induced Pluripotent Stem Cells/drug effects
• Neurites/drug effects
• Neurites/metabolism
• Neurons/cytology
• Neurons/drug effects
• Neurons/physiology
• Protein Kinase Inhibitors/pharmacology
• Pyridines/pharmacology
• rho-Associated Kinases/antagonists & inhibitors
• rho-Associated Kinases/metabolism

• T32 GM007267 NIGMS NIH HHS
• R01NS099586-01 NINDS NIH HHS
• T32 GM008136 NIGMS NIH HHS
• U01 MH106882 NIMH NIH HHS
• T32GM008328-24 NIGMS NIH HHS
• R01 NS099586 NINDS NIH HHS
• T32 GM008328 NIGMS NIH HHS

Adult human multipotent neural cell (ahMNC) is a candidate for regeneration therapy for neurodegenerative diseases. Here, we developed a primary clump culture method for ahMNCs to increase the efficiency of isolation and in vitro expansion. The same amount of human temporal lobe (1 g) was partially digested and then filtered through strainers with various pore sizes, resulting in four types of clumps: Clump I > 100 µm, 70 µm < Clump II < 100 µm, 40 µm < Clump III < 70 µm, and Clump IV < 40 µm. At 3 and 6 days after culture, Clump II showed significantly higher number of colonies than the other Clumps. Moreover, ahMNCs derived from Clump II (ahMNCs-Clump II) showed stable proliferation, and shortened the time to first passage from 19 to 15 days, and the time to 1 × 10⁹ cells from 42 to 34 days compared with the previous single-cell method. ahMNCs-Clump II had neural differentiation and pro-angiogenic potentials, which are the characteristics of ahMNCs. In conclusion, the novel clump culture method for ahMNCs has significantly higher efficiency than previous techniques. Considering the small amount of available human brain tissue, the clump culture method would promote further clinical applications of ahMNCs.

• adult human multipotent neural cells
• angiogenic potential
• clump culture
• neural differentiation

• Adult
• Adult Stem Cells/cytology
• Cell Culture Techniques/methods
• Cell Differentiation
• Cell Proliferation
• Cells, Cultured
• Human Umbilical Vein Endothelial Cells
• Humans
• Multipotent Stem Cells/cytology
• Neovascularization, Physiologic
• Neural Stem Cells/cytology

• Epilepsy
• Human neural stem cells
• Inflammation
• Tissue engineering
• Traumatic brain injury

Direct-lineage conversion of the somatic cell by reprogramming, in which mature cells were fully converted into a variety of other cell types bypassing an intermediate pluripotent state, is a promising regenerative medicine approach. Due to the risk of tumorigenesis by viral methods, a non-viral carrier for the delivery of reprogramming factors is very desirable. This study utilized the mesoporous silica nanoparticles (MSNs) as a non-viral delivery system for transduction of the three key factors to achieve conversion of mouse fibroblasts (MFs) into functional dopaminergic neuron-like cells (denoted as fDA-neurons). At the same time, a neurogenesis inducer, ISX-9, was co-delivered with the MSNs to promote the direct conversion of neuron-like cells. Good transfection efficiency of plasmid@MSN allowed repeated dosing to maintain high exogenous gene expression analyzed by qPCR and the changes in neural function markers were monitored. To further validate the dopaminergic function and the electrophysiological properties of fDA-neurons, the results of ELISA assay showed the high levels of secreted-dopamine in the conditional medium and rich Na⁺/K⁺-channels were observed in the fDA-neurons on Day 22. The results demonstrated that MSN nanocarrier is effective in delivering the reprogramming factors for the conversion of functional dopaminergic neurons from adult somatic cells.

• Müller glia
• Neural
• differentiation
• peripheral retina
• progenitor cell
• stem cell

The adult brain is unable to regenerate itself sufficiently after large injuries. Therefore, hopes rely on therapies using neural stem cell or biomaterial transplantation to sustain brain reconstruction. The aim of the present study was to evaluate the improvement in sensorimotor recovery brought about by human primary adult neural stem cells (hNSCs) in combination with bio-implants.

hNSCs were pre-seeded on implants micropatterned for neurite guidance and inserted intracerebrally 2 weeks after a primary motor cortex lesion in rats. Long-term behaviour was significantly improved after hNSC implants versus cell engraftment in the grip strength test. MRI and immunohistological studies were conducted to elucidate the underlying mechanisms of neuro-implant integration.

hNSC implants promoted tissue reconstruction and limited hemispheric atrophy and glial scar expansion. After 3 months, grafted hNSCs were detected on implants and expressed mature neuronal markers (NeuN, MAP2, SMI312). They also migrated over a short distance to the reconstructed tissues and to the peri-lesional tissues, where 26% integrated as mature neurons. Newly formed host neural progenitors (nestin, DCX) colonized the implants, notably in the presence of hNSCs, and participated in tissue reconstruction. The microstructured bio-implants sustained the guided maturation of both grafted hNSCs and endogenous progenitors.

These immunohistological results are coherent with and could explain the late improvement observed in sensorimotor recovery. These findings provide novel insights into the regenerative potential of primary adult hNSCs combined with microstructured implants.

• Biomaterial
• Brain injury
• Cell therapy
• Sensorimotor recovery
• Tissue engineering

• Human fetal neural stem cell
• Neurogenesis
• Optoelectrical stimulation
• Photoactive nanoweb substrate
• Topographical stimulation

In our lab we study neurogenesis and the development of brain tumors. We work towards treatment strategies for glioblastoma and towards using autologous neural stem cells for tissue regeneration strategies for brain damage and neurodegenerative disorders. It has been our policy to try to establish living cell cultures from all human biopsy material that we obtain. We hypothesized that small pieces of brain tissue could be cryopreserved and that live neural stem cells could be recovered at a later time. DMSO has been shown to possess a remarkable ability to diffuse through cell membranes and pass into cell interiors. Its chemical properties prevent the formation of damaging ice crystals thus allowing cell storage at or below −180 C. We report here a protocol for successful freezing of small pieces of tissue derived from human brain and human brain tumours. Virtually all specimens could be successfully revived. Assays of phenotype and behaviour show that the cell cultures derived were equivalent to those cultures previously derived from fresh tissue.

Glioblastoma (GBM) is both the most common and the most lethal primary brain tumor. It is thought that GBM stem cells (GSCs) are critically important in resistance to therapy. Therefore, there is a strong rationale to target these cells in order to develop new molecular therapies.

To identify molecular targets in GSCs, we compared gene expression in GSCs to that in neural stem cells (NSCs) from the adult human brain, using microarrays. Bioinformatic filtering identified 20 genes (PBK/TOPK, CENPA, KIF15, DEPDC1, CDC6, DLG7/DLGAP5/HURP, KIF18A, EZH2, HMMR/RHAMM/CD168, NOL4, MPP6, MDM1, RAPGEF4, RHBDD1, FNDC3B, FILIP1L, MCC, ATXN7L4/ATXN7L1, P2RY5/LPAR6 and FAM118A) that were consistently expressed in GSC cultures and consistently not expressed in NSC cultures. The expression of these genes was confirmed in clinical samples (TCGA and REMBRANDT). The first nine genes were highly co-expressed in all GBM subtypes and were part of the same protein-protein interaction network. Furthermore, their combined up-regulation correlated negatively with patient survival in the mesenchymal GBM subtype. Using targeted proteomics and the COGNOSCENTE database we linked these genes to GBM signalling pathways.

Nine genes: PBK, CENPA, KIF15, DEPDC1, CDC6, DLG7, KIF18A, EZH2 and HMMR should be further explored as targets for treatment of GBM.

• GBM
• GSCs
• glioblastoma
• glioblastoma stem cells
• therapeutic targeting

• Electrophysiology
• Human striatal primordium
• Neurotrophic factor BDNF
• Neurotrophic factor FGF2

Stem cell-based therapy is a potential treatment for neurodegenerative diseases, but its application to Alzheimer’s disease (AD) remains limited. Brain-derived neurotrophic factor (BDNF) is critical in the pathogenesis and treatment of AD. Here, we present a novel therapeutic approach for AD treatment using BDNF-overexpressing neural stem cells (BDNF-NSCs). In vitro, BDNF overexpression was neuroprotective to beta-amyloid-treated NSCs. In vivo, engrafted BDNF-NSCs-derived neurons not only displayed the Ca²⁺-response fluctuations, exhibited electrophysiological properties of mature neurons and integrated into local brain circuits, but recovered the cognitive deficits. Furthermore, BDNF overexpression improved the engrafted cells’ viability, neuronal fate, neurite complexity, maturation of electrical property and the synaptic density. In contrast, knockdown of the BDNF in BDNF-NSCs diminished stem cell-based therapeutic efficacy. Together, our findings indicate BDNF overexpression improves the therapeutic potential of engrafted NSCs for AD via neurogenic effects and neuronal replacement, and further support the feasibility of NSC-based ex vivo gene therapy for AD.

• Cell replacement therapy
• Enteric nervous system
• Enteric neuropathies
• Hirschsprung disease
• Stem cells

Over the past two decades, regenerative therapies using stem cell technologies have been developed for various neurological diseases. Although stem cell therapy is an attractive option to reverse neural tissue damage and to recover neurological deficits, it is still under development so as not to show significant treatment effects in clinical settings. In this review, we discuss the scientific and clinical basics of adult neural stem cells (aNSCs), and their current developmental status as cell therapeutics for neurological disease. Compared with other types of stem cells, aNSCs have clinical advantages, such as limited proliferation, inborn differentiation potential into functional neural cells, and no ethical issues. In spite of the merits of aNSCs, difficulties in the isolation from the normal brain, and in the in vitro expansion, have blocked preclinical and clinical study using aNSCs. However, several groups have recently developed novel techniques to isolate and expand aNSCs from normal adult brains, and showed successful applications of aNSCs to neurological diseases. With new technologies for aNSCs and their clinical strengths, previous hurdles in stem cell therapies for neurological diseases could be overcome, to realize clinically efficacious regenerative stem cell therapeutics.

• Adult neural stem cell
• Neurological diseases
• Stem cell therapy
• Preclinical trial
• Clinical trial

• CRAC channels
• Orai1
• STIM1
• calcium
• progenitor cell
• proliferation

Many protocols have been designed to differentiate human embryonic stem cells (ESCs) and human induced pluripotent stem cells (iPSCs) into neurons. Despite the relevance of electrophysiological properties for proper neuronal function, little is known about the evolution over time of important neuronal electrophysiological parameters in iPSC-derived neurons. Yet, understanding the development of basic electrophysiological characteristics of iPSC-derived neurons is critical for evaluating their usefulness in basic and translational research. Therefore, we analyzed the basic electrophysiological parameters of forebrain neurons differentiated from human iPSCs, from day 31 to day 55 after the initiation of neuronal differentiation. We assayed the developmental progression of various properties, including resting membrane potential, action potential, sodium and potassium channel currents, somatic calcium transients and synaptic activity. During the maturation of iPSC-derived neurons, the resting membrane potential became more negative, the expression of voltage-gated sodium channels increased, the membrane became capable of generating action potentials following adequate depolarization and, at day 48–55, 50% of the cells were capable of firing action potentials in response to a prolonged depolarizing current step, of which 30% produced multiple action potentials. The percentage of cells exhibiting miniature excitatory post-synaptic currents increased over time with a significant increase in their frequency and amplitude. These changes were associated with an increase of Ca²⁺ transient frequency. Co-culturing iPSC-derived neurons with mouse glial cells enhanced the development of electrophysiological parameters as compared to pure iPSC-derived neuronal cultures. This study demonstrates the importance of properly evaluating the electrophysiological status of the newly generated neurons when using stem cell technology, as electrophysiological properties of iPSC-derived neurons mature over time.

Prevailingly, adult mammalian neurogenesis is thought to occur in discrete, separate locations known as neurogenic niches that are best characterized in the subgranular zone (SGZ) of the dentate gyrus and in the subventricular zone (SVZ). The existence of adult human neurogenic niches is controversial.

The existence of neurogenic niches was investigated with neurogenesis marker immunostaining in histologically normal human brains obtained from autopsies. Twenty-eight adult temporal lobes, specimens from limbic structures and the hypothalamus of one newborn and one adult were examined.

The neural stem cell marker nestin stained circumventricular organ cells and the immature neuronal marker doublecortin (DCX) stained hypothalamic and limbic structures adjacent to circumventricular organs; both markers stained a continuous structure running from the hypothalamus to the hippocampus. The cell proliferation marker Ki-67 was detected predominately in structures that form the septo-hypothalamic continuum. Nestin-expressing cells were located in the fimbria-fornix at the insertion of the choroid plexus; ependymal cells in this structure expressed the putative neural stem cell marker CD133. From the choroidal fissure in the temporal lobe, a nestin-positive cell layer spread throughout the SVZ and subpial zone. In the subpial zone, a branch of this layer reached the hippocampal sulcus and ended in the SGZ (principally in the newborn) and in the subiculum (principally in the adults). Another branch of the nestin-positive cell layer in the subpial zone returned to the optic chiasm. DCX staining was detected in the periventricular and middle hypothalamus and more densely from the mammillary body to the subiculum through the fimbria-fornix, thus running through the principal neuronal pathway from the hippocampus to the hypothalamus. The column of the fornix forms part of this pathway and appears to coincide with the zone previously identified as the human rostral migratory stream. Partial co-labeling with DCX and the neuronal marker βIII-tubulin was also observed.

Collectively, these findings suggest the existence of an adult human neurogenic system that rises from the circumventricular organs and follows, at minimum, the circuitry of the hypothalamus and limbic system.

The growth and recurrence of several cancers appear to be driven by a population of cancer stem cells (CSCs). Glioblastoma, the most common primary brain tumor, is invariably fatal, with a median survival of approximately 1 year. Although experimental data have suggested the importance of CSCs, few data exist regarding the potential relevance and importance of these cells in a clinical setting.

We here present the first seven patients treated with a dendritic cell (DC)-based vaccine targeting CSCs in a solid tumor. Brain tumor biopsies were dissociated into single-cell suspensions, and autologous CSCs were expanded in vitro as tumorspheres. From these, CSC-mRNA was amplified and transfected into monocyte-derived autologous DCs. The DCs were aliquoted to 9–18 vaccines containing 10⁷ cells each. These vaccines were injected intradermally at specified intervals after the patients had received a standard 6-week course of post-operative radio-chemotherapy. The study was registered with the ClinicalTrials.gov identifier NCT00846456.

Autologous CSC cultures were established from ten out of eleven tumors. High-quality RNA was isolated, and mRNA was amplified in all cases. Seven patients were able to be weaned from corticosteroids to receive DC immunotherapy. An immune response induced by vaccination was identified in all seven patients. No patients developed adverse autoimmune events or other side effects. Compared to matched controls, progression-free survival was 2.9 times longer in vaccinated patients (median 694 vs. 236 days, p = 0.0018, log-rank test).

These findings suggest that vaccination against glioblastoma stem cells is safe, well-tolerated, and may prolong progression-free survival.

The online version of this article (doi:10.1007/s00262-013-1453-3) contains supplementary material, which is available to authorized users.

• Adult human neural stem cell
• CSC
• Cancer stem cell
• ESC
• Embryonic stem cell
• GBM
• GSC
• Glioblastoma
• Glioma stem cell
• HSC
• Inducible pluripotent stem cell
• LSC
• Leukemic stem cell
• MSigDB
• Molecular Signature Database
• Sfrp1
• Survival
• Wnt
• ahNSC
• hematopoetic stem cell
• iPSC

• Cell Differentiation/genetics
• Cell Differentiation/physiology
• Cells, Cultured
• Electrophysiology
• Epithelial Cells/cytology
• Epithelial Cells/metabolism
• Humans
• Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
• Motor Neurons/cytology
• Motor Neurons/metabolism
• Neural Crest/cytology
• Neural Crest/metabolism
• Neural Stem Cells/cytology
• Neural Stem Cells/metabolism
• Neurodegenerative Diseases/genetics
• Neurodegenerative Diseases/metabolism
• Neurons/cytology
• Neurons/metabolism
• Protein Serine-Threonine Kinases/genetics
• Protein Serine-Threonine Kinases/metabolism

• Enteric nervous system
• Neural activity development
• Neuron differentiation
• Synapse formation

The ability to culture neural progenitor cells from the adult human brain has provided an exciting opportunity to develop and test potential therapies on adult human brain cells. To achieve a reliable and reproducible adult human neural progenitor cell (AhNPC) culture system for this purpose, this study fully characterized the cellular composition of the AhNPC cultures, as well as the possible changes to this in vitro system over prolonged culture periods. We isolated cells from the neurogenic subventricular zone/hippocampus (SVZ/HP) of the adult human brain and found a heterogeneous culture population comprised of several types of post-mitotic brain cells (neurons, astrocytes, and microglia), and more importantly, two distinct mitotic cell populations; the AhNPCs, and the fibroblast-like cells (FbCs). These two populations can easily be mistaken for a single population of AhNPCs, as they both proliferate under AhNPC culture conditions, form spheres and express neural progenitor cell and early neuronal markers, all of which are characteristics of AhNPCs in vitro. However, despite these similarities under proliferating conditions, under neuronal differentiation conditions, only the AhNPCs differentiated into functional neurons and glia. Furthermore, AhNPCs showed limited proliferative capacity that resulted in their depletion from culture by 5–6 passages, while the FbCs, which appear to be from a neurovascular origin, displayed a greater proliferative capacity and dominated the long-term cultures. This gradual change in cellular composition resulted in a progressive decline in neurogenic potential without the apparent loss of self-renewal in our cultures. These results demonstrate that while AhNPCs and FbCs behave similarly under proliferative conditions, they are two different cell populations. This information is vital for the interpretation and reproducibility of AhNPC experiments and suggests an ideal time frame for conducting AhNPC-based experiments.

• Adult
• Biopsy
• Brain/cytology
• Cell Differentiation/drug effects
• Cell Separation
• Fibroblasts/cytology
• Fibroblasts/drug effects
• Fibroblasts/metabolism
• Humans
• Laminin/pharmacology
• Lung/cytology
• Mitosis/drug effects
• Neovascularization, Physiologic/drug effects
• Neural Stem Cells/cytology
• Neural Stem Cells/drug effects
• Neural Stem Cells/metabolism
• Neurogenesis/drug effects
• Neurons/cytology
• Neurons/drug effects
• Neurons/metabolism

• Adolescent
• Adult
• Aged
• Aged, 80 and over
• Biomarkers/metabolism
• Cell Proliferation
• Cell Separation
• Cells, Cultured
• Ciliary Body/cytology
• Ciliary Body/metabolism
• DNA Primers/chemistry
• Fluorescent Antibody Technique, Indirect
• Glaucoma/surgery
• Humans
• Iridectomy
• Iris/cytology
• Iris/metabolism
• Microscopy, Electron, Transmission
• Middle Aged
• Pigment Epithelium of Eye/cytology
• Pigment Epithelium of Eye/metabolism
• Retinal Pigment Epithelium/cytology
• Retinal Pigment Epithelium/metabolism
• Reverse Transcriptase Polymerase Chain Reaction
• Stem Cells/metabolism
• Stem Cells/ultrastructure

Filum terminale (FT) is a structure that is intimately associated with conus medullaris, the most caudal part of the spinal cord. It is well documented that certain regions of the adult human central nervous system contains undifferentiated, progenitor cells or multipotent precursors. The primary objective of this study was to describe the distribution and progenitor features of this cell population in humans, and to confirm their ability to differentiate within the neuroectodermal lineage.

We demonstrate that neural stem/progenitor cells are present in FT obtained from patients treated for tethered cord. When human or rat FT-derived cells were cultured in defined medium, they proliferated and formed neurospheres in 13 out of 21 individuals. Cells expressing Sox2 and Musashi-1 were found to outline the central canal, and also to be distributed in islets throughout the whole FT. Following plating, the cells developed antigen profiles characteristic of astrocytes (GFAP) and neurons (β-III-tubulin). Addition of PDGF-BB directed the cells towards a neuronal fate. Moreover, the cells obtained from young donors shows higher capacity for proliferation and are easier to expand than cells derived from older donors.

The identification of bona fide neural progenitor cells in FT suggests a possible role for progenitor cells in this extension of conus medullaris and may provide an additional source of such cells for possible therapeutic purposes.

Filum terminale, human, progenitor cells, neuron, astrocytes, spinal cord.