1,707 results on '"Neurons transplantation"'
Search Results
2. Hybrid brains: the ethics of transplanting human neurons into animals.
- Author
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Powell K
- Subjects
- Animals, Humans, Animal Experimentation ethics, Brain cytology, Cell Transplantation ethics, Ethics, Research, Heterografts cytology, Neurons transplantation, Transplantation, Heterologous ethics
- Published
- 2022
- Full Text
- View/download PDF
3. LRP10 interacts with SORL1 in the intracellular vesicle trafficking pathway in non-neuronal brain cells and localises to Lewy bodies in Parkinson's disease and dementia with Lewy bodies.
- Author
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Grochowska MM, Carreras Mascaro A, Boumeester V, Natale D, Breedveld GJ, Geut H, van Cappellen WA, Boon AJW, Kievit AJA, Sammler E, Parchi P, Cortelli P, Alessi DR, van de Berg WDJ, Bonifati V, and Mandemakers W
- Subjects
- Adult, Aged, Astrocytes metabolism, Astrocytes transplantation, Brain cytology, Brain pathology, Genetic Variation, Humans, Induced Pluripotent Stem Cells transplantation, Lewy Bodies pathology, Lewy Body Disease pathology, Middle Aged, Neurodegenerative Diseases pathology, Neurons transplantation, Parkinson Disease pathology, Brain metabolism, LDL-Receptor Related Proteins genetics, Lewy Bodies metabolism, Lewy Body Disease metabolism, Membrane Transport Proteins genetics, Parkinson Disease metabolism
- Abstract
Loss-of-function variants in the low-density lipoprotein receptor-related protein 10 (LRP10) gene have been associated with autosomal-dominant Parkinson's disease (PD), PD dementia, and dementia with Lewy bodies (DLB). Moreover, LRP10 variants have been found in individuals diagnosed with progressive supranuclear palsy and amyotrophic lateral sclerosis. Despite this genetic evidence, little is known about the expression and function of LRP10 protein in the human brain under physiological or pathological conditions. To better understand how LRP10 variants lead to neurodegeneration, we first performed an in-depth characterisation of LRP10 expression in post-mortem brains and human-induced pluripotent stem cell (iPSC)-derived astrocytes and neurons from control subjects. In adult human brain, LRP10 is mainly expressed in astrocytes and neurovasculature but undetectable in neurons. Similarly, LRP10 is highly expressed in iPSC-derived astrocytes but cannot be observed in iPSC-derived neurons. In astrocytes, LRP10 is present at trans-Golgi network, plasma membrane, retromer, and early endosomes. Interestingly, LRP10 also partially co-localises and interacts with sortilin-related receptor 1 (SORL1). Furthermore, although LRP10 expression and localisation in the substantia nigra of most idiopathic PD and DLB patients and LRP10 variant carriers diagnosed with PD or DLB appeared unchanged compared to control subjects, significantly enlarged LRP10-positive vesicles were detected in a patient carrying the LRP10 p.Arg235Cys variant. Last, LRP10 was detected in Lewy bodies (LB) at late maturation stages in brains from idiopathic PD and DLB patients and in LRP10 variant carriers. In conclusion, high LRP10 expression in non-neuronal cells and undetectable levels in neurons of control subjects indicate that LRP10-mediated pathogenicity is initiated via cell non-autonomous mechanisms, potentially involving the interaction of LRP10 with SORL1 in vesicle trafficking pathways. Together with the specific pattern of LRP10 incorporation into mature LBs, these data support an important mechanistic role for disturbed vesicle trafficking and loss of LRP10 function in neurodegenerative diseases.
- Published
- 2021
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4. Cellular complexity in brain organoids: Current progress and unsolved issues.
- Author
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Mansour AA, Schafer ST, and Gage FH
- Subjects
- Brain physiology, Cell Differentiation, Cell Transplantation methods, Cell Transplantation trends, Endothelial Cells cytology, Endothelial Cells physiology, Humans, Lymphocytes cytology, Lymphocytes physiology, Models, Biological, Neovascularization, Physiologic, Neural Stem Cells physiology, Neural Stem Cells transplantation, Neurogenesis physiology, Neuroglia cytology, Neuroglia physiology, Neurons physiology, Neurons transplantation, Organoids physiology, Pluripotent Stem Cells physiology, Brain cytology, Neural Stem Cells cytology, Neurons cytology, Organoids cytology, Pluripotent Stem Cells cytology
- Abstract
Brain organoids are three-dimensional neural aggregates derived from pluripotent stem cells through self-organization and recapitulate architectural and cellular aspects of certain brain regions. Brain organoids are currently a highly exciting area of research that includes the study of human brain development, function, and dysfunction in unprecedented ways. In this Review, we discuss recent discoveries related to the generation of brain organoids that resemble diverse brain regions. We provide an overview of the strategies to complement these primarily neuroectodermal models with cell types of non-neuronal origin, such as vasculature and immune cells. Recent transplantation approaches aiming to achieve higher cellular complexity and long-term survival of these models will then be discussed. We conclude by highlighting unresolved key questions and future directions in this exciting area of human brain organogenesis., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2021
- Full Text
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5. Bioengineering tissue morphogenesis and function in human neural organoids.
- Author
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Fedorchak NJ, Iyer N, and Ashton RS
- Subjects
- Bioengineering methods, Brain physiology, Cell Differentiation, Endothelial Cells cytology, Endothelial Cells physiology, Extracellular Matrix metabolism, Humans, Models, Biological, Neovascularization, Physiologic, Neural Stem Cells physiology, Neural Stem Cells transplantation, Neurogenesis physiology, Neuroglia cytology, Neuroglia physiology, Neurons physiology, Neurons transplantation, Organoids physiology, Pluripotent Stem Cells physiology, Tissue Engineering methods, Brain cytology, Morphogenesis physiology, Neural Stem Cells cytology, Neurons cytology, Organoids cytology, Pluripotent Stem Cells cytology
- Abstract
Over the last decade, scientists have begun to model CNS development, function, and disease in vitro using human pluripotent stem cell (hPSC)-derived organoids. Using traditional protocols, these 3D tissues are generated by combining the innate emergent properties of differentiating hPSC aggregates with a bioreactor environment that induces interstitial transport of oxygen and nutrients and an optional supportive hydrogel extracellular matrix (ECM). During extended culture, the hPSC-derived neural organoids (hNOs) obtain millimeter scale sizes with internal microscale cytoarchitectures, cellular phenotypes, and neuronal circuit behaviors mimetic of those observed in the developing brain, eye, or spinal cord. Early studies evaluated the cytoarchitectural and phenotypical character of these organoids and provided unprecedented insight into the morphogenetic processes that govern CNS development. Comparisons to human fetal tissues revealed their significant similarities and differences. While hNOs have current disease modeling applications and significant future promise, their value as anatomical and physiological models is limited because they fail to form reproducibly and recapitulate more mature in vivo features. These include biomimetic macroscale tissue morphology, positioning of morphogen signaling centers to orchestrate appropriate spatial organization and intra- and inter-connectivity of discrete tissue regions, maturation of physiologically relevant neural circuits, and formation of vascular networks that can support sustained in vitro tissue growth. To address these inadequacies scientists have begun to integrate organoid culture with bioengineering techniques and methodologies including genome editing, biomaterials, and microfabricated and microfluidic platforms that enable spatiotemporal control of cellular differentiation or the biochemical and biophysical cues that orchestrate organoid morphogenesis. This review will examine recent advances in hNO technologies and culture strategies that promote reproducible in vitro morphogenesis and greater biomimicry in structure and function., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2021
- Full Text
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6. Lipocalin-2 mediates the rejection of neural transplants.
- Author
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Weng YC, Huang YT, Chiang IC, Tsai PJ, Su YW, and Chou WH
- Subjects
- Animals, Apoptosis genetics, Apoptosis physiology, Brain cytology, Brain metabolism, Cells, Cultured, Flow Cytometry, Graft Rejection genetics, Humans, Immunohistochemistry, In Situ Nick-End Labeling, Lipocalin-2 genetics, Male, Mice, Mice, Inbred C57BL, Real-Time Polymerase Chain Reaction, Graft Rejection metabolism, Lipocalin-2 metabolism, Lipocalin-2 physiology, Neurons metabolism, Neurons transplantation
- Abstract
Lipocalin-2 (LCN2) has been implicated in promoting apoptosis and neuroinflammation in neurological disorders; however, its role in neural transplantation remains unknown. In this study, we cultured and differentiated Lund human mesencephalic (LUHMES) cells into human dopaminergic-like neurons and found that LCN2 mRNA was progressively induced in mouse brain after the intrastriatal transplantation of human dopaminergic-like neurons. The induction of LCN2 protein was detected in a subset of astrocytes and neutrophils infiltrating the core of the engrafted sites, but not in neurons and microglia. LCN2-immunoreactive astrocytes within the engrafted sites expressed lower levels of A1 and A2 astrocytic markers. Recruitment of microglia, neutrophils, and monocytes after transplantation was attenuated in LCN2 deficiency mice. The expression of M2 microglial markers was significantly elevated and survival of engrafted neurons was markedly improved after transplantation in LCN2 deficiency mice. Brain type organic cation transporter (BOCT), the cell surface receptor for LCN2, was induced in dopaminergic-like neurons after differentiation, and treatment with recombinant LCN2 protein directly induced apoptosis in dopaminergic-like neurons in a dose-dependent manner. Our results, therefore, suggested that LCN2 is a neurotoxic factor for the engrafted neurons and a modulator of neuroinflammation. LCN2 inhibition may be useful in reducing rejection after neural transplantation., (© 2020 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)
- Published
- 2021
- Full Text
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7. Immune-tolerance to human iPS-derived neural progenitors xenografted into the immature cerebellum is overridden by species-specific differences in differentiation timing.
- Author
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Nato G, Corti A, Parmigiani E, Jachetti E, Lecis D, Colombo MP, Delia D, Buffo A, and Magrassi L
- Subjects
- Animals, Cells, Cultured, Cerebellum growth & development, Female, Humans, Mice, Mice, Inbred NOD, Mice, SCID, Neurons cytology, Rats, Rats, Wistar, Species Specificity, Transplantation, Heterologous, Cell Differentiation, Cerebellum immunology, Graft Survival, Induced Pluripotent Stem Cells cytology, Neural Stem Cells cytology, Neurons transplantation, Stem Cell Transplantation methods
- Abstract
We xeno-transplanted human neural precursor cells derived from induced pluripotent stem cells into the cerebellum and brainstem of mice and rats during prenatal development or the first postnatal week. The transplants survived and started to differentiate up to 1 month after birth when they were rejected by both species. Extended survival and differentiation of the same cells were obtained only when they were transplanted in NOD-SCID mice. Transplants of human neural precursor cells mixed with the same cells after partial in vitro differentiation or with a cellular extract obtained from adult rat cerebellum increased survival of the xeno-graft beyond one month. These findings are consistent with the hypothesis that the slower pace of differentiation of human neural precursors compared to that of rodents restricts induction of immune-tolerance to human antigens expressed before completion of maturation of the immune system. With further maturation the transplanted neural precursors expressed more mature antigens before the graft were rejected. Supplementation of the immature cells suspensions with more mature antigens may help to induce immune-tolerance for those antigens expressed only later by the engrafted cells.
- Published
- 2021
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8. Preparation of Neural Stem Cells and Progenitors: Neuronal Production and Grafting Applications.
- Author
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Zholudeva LV, Jin Y, Qiang L, Lane MA, and Fischer I
- Subjects
- Animals, Cell Culture Techniques, Cell Lineage, Cell Proliferation, Cell Separation, Cell Survival, Cells, Cultured, Cellular Reprogramming, Cellular Reprogramming Techniques, Female, Gene Expression Regulation, Developmental, Gestational Age, Humans, Induced Pluripotent Stem Cells physiology, Mice, Neural Stem Cells physiology, Neurons physiology, Phenotype, Pregnancy, Rats, Induced Pluripotent Stem Cells transplantation, Neural Stem Cells transplantation, Neurogenesis, Neurons transplantation, Spinal Cord embryology
- Abstract
Neural stem cells (NSCs) are a valuable tool for the study of neural development and function as well as an important source of cell transplantation strategies for neural disease. NSCs can be used to study how neurons acquire distinct phenotypes and how the interactions between neurons and glial cells in the developing nervous system shape the structure and function of the CNS. NSCs can also be used for cell replacement therapies following CNS injury targeting astrocytes, oligodendrocytes, and neurons. With the availability of patient-derived induced pluripotent stem cells (iPSCs), neurons prepared from NSCs can be used to elucidate the molecular basis of neurological disorders leading to potential treatments. Although NSCs can be derived from different species and many sources, including embryonic stem cells (ESCs), iPSCs, adult CNS, and direct reprogramming of nonneural cells, isolating primary NSCs directly from fetal tissue is still the most common technique for preparation and study of neurons. Regardless of the source of tissue, similar techniques are used to maintain NSCs in culture and to differentiate NSCs toward mature neural lineages. This chapter will describe specific methods for isolating and characterizing multipotent NSCs and neural precursor cells (NPCs) from embryonic rat CNS tissue (mostly spinal cord) and from human ESCs and iPSCs as well as NPCs prepared by reprogramming. NPCs can be separated into neuronal and glial restricted progenitors (NRP and GRP, respectively) and used to reliably produce neurons or glial cells both in vitro and following transplantation into the adult CNS. This chapter will describe in detail the methods required for the isolation, propagation, storage, and differentiation of NSCs and NPCs isolated from rat and mouse spinal cords for subsequent in vitro or in vivo studies as well as new methods associated with ESCs, iPSCs, and reprogramming.
- Published
- 2021
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9. Glial cells in Parkinson´s disease: protective or deleterious?
- Author
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Domingues AV, Pereira IM, Vilaça-Faria H, Salgado AJ, Rodrigues AJ, and Teixeira FG
- Subjects
- Central Nervous System pathology, Humans, Nerve Degeneration pathology, Nerve Degeneration therapy, Neurons pathology, Parkinson Disease pathology, Cell- and Tissue-Based Therapy, Neuroglia transplantation, Neurons transplantation, Parkinson Disease therapy
- Abstract
Glial cells have been identified more than 100 years ago, and are known to play a key role in the central nervous system (CNS) function. A recent piece of evidence is emerging showing that in addition to the capacity of CNS modulation and homeostasis, glial cells are also being looked like as a promising cell source not only to study CNS pathologies initiation and progression but also to the establishment and development of new therapeutic strategies. Thus, in the present review, we will discuss the current evidence regarding glial cells' contribution to neurodegenerative diseases as Parkinson's disease, providing cellular, molecular, functional, and behavioral data supporting its active role in disease initiation, progression, and treatment. As so, considering their functional relevance, glial cells may be important to the understanding of the underlying mechanisms regarding neuronal-glial networks in neurodegeneration/regeneration processes, which may open new research opportunities for their future use as a target or treatment in human clinical trials.
- Published
- 2020
- Full Text
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10. Orexin cell transplant reduces behavioral arrest severity in narcoleptic mice.
- Author
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Equihua-Benítez AC, Equihua-Benítez JA, Guzmán-Vásquez K, Prospero-García O, and Drucker-Colín R
- Subjects
- Animals, Disease Models, Animal, Hypothalamus cytology, Hypothalamus metabolism, Mice, Mice, Transgenic, Neurons metabolism, Narcolepsy, Neurons transplantation, Orexins metabolism
- Abstract
Narcolepsy is a sleep disorder that has been associated with the loss of orexinergic neurons from the lateral hypothalamic area. This loss leads to dysregulated sleep and cataplexy attacks. Therapeutic options are currently limited to symptom management with pharmacotherapy and nonpharmacological approaches. Nonetheless, cell replacement therapy could offer relief, and research in the field has yielded positive results for other neurodegenerative disorders, such as Parkinson's disease. Thus, we propose that orexin cell rich grafts could help improve narcoleptic symptoms in the orexin/ataxin-3 mouse model of narcolepsy. For this purpose, we isolated EGFP+ cells from either orexin/EGFP or CAG-EGFP mice with the use of a flow cytometer and grafted them into the pedunculopontine and laterodorsal tegmentum nuclei (PPT/LDDT) of orexin/ataxin-3 mice. Our results show that even small orexinergic grafts can reduce the severity of behavioral arrests, with a median reduction of 30.31% in episode duration, 51.35% for number of events and 69.73% in time spent in the behavioral arrest state and help with sleep fragmentation measured in number of bouts per behavioral state. Surprisingly, control grafts made from cerebellar tissue also reduced behavioral arrest severity, but to a lesser degree. Although still at a very early stage, these results show that there is potential in cell grafts for improving aspects of the narcoleptic phenotype and further research could help elucidate realistic expectations of an orexin cell replacement therapy for narcolepsy., (Copyright © 2020. Published by Elsevier B.V.)
- Published
- 2020
- Full Text
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11. The Fate of Transplanted Olfactory Progenitors Is Conditioned by the Cell Phenotypes of the Receiver Brain Tissue in Cocultures.
- Author
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Pourié G, Akchiche N, Millot JL, Guéant JL, Daval JL, and Bossenmeyer-Pourié C
- Subjects
- Animals, Brain cytology, Brain growth & development, Cell Differentiation genetics, Cell Lineage genetics, Central Nervous System metabolism, Coculture Techniques, Humans, Mice, Nerve Growth Factor genetics, Neuroglia cytology, Neuroglia metabolism, Neuroglia transplantation, Neurons transplantation, Olfactory Cortex cytology, Olfactory Cortex transplantation, Oligodendroglia cytology, Oligodendroglia metabolism, Oligodendroglia transplantation, Stem Cells metabolism, Brain metabolism, Neurons metabolism, Olfactory Cortex metabolism, Stem Cell Transplantation, Stem Cells cytology
- Abstract
Among the numerous candidates for cell therapy of the central nervous system (CNS), olfactory progenitors (OPs) represent an interesting alternative because they are free of ethical concerns, are easy to collect, and allow autologous transplantation. In the present study, we focused on the optimization of neuron production and maturation. It is known that plated OPs respond to various trophic factors, and we also showed that the use of Nerve Growth Factor (NGF) allowed switching from a 60/40 neuron/glia ratio to an 80/20 one. Nevertheless, in order to focus on the integration of OPs in mature neural circuits, we cocultured OPs in primary cultures obtained from the cortex and hippocampus of newborn mice. When dissociated OPs were plated, they differentiated into both glial and neuronal phenotypes, but we obtained a 1.5-fold higher viability in cortex/OP cocultures than in hippocampus/OP ones. The fate of OPs in cocultures was characterized with different markers such as BrdU, Map-2, and Synapsin, indicating a healthy integration. These results suggest that the integration of transplanted OPs might by affected by trophic factors and the environmental conditions/cell phenotypes of the host tissue. Thus, a model of coculture could provide useful information on key cell events for the use of progenitors in cell therapy.
- Published
- 2020
- Full Text
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12. Stem Cell-Based Disease Modeling and Cell Therapy.
- Author
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Bai X
- Subjects
- Gene Editing methods, Humans, Neurons pathology, Neurons transplantation, Organoids transplantation, Stem Cell Research, CRISPR-Cas Systems genetics, Cell Differentiation genetics, Cell- and Tissue-Based Therapy, Induced Pluripotent Stem Cells transplantation
- Abstract
Stem cell science is among the fastest moving fields in biology, with many highly promising directions for translatability. To centralize and contextualize some of the latest developments, this Special Issue presents state-of-the-art research of adult stem cells, induced pluripotent stem cells (iPSCs), and embryonic stem cells as well as cancer stem cells. The studies we include describe efficient differentiation protocols of generation of chondrocytes, adipocytes, and neurons, maturation of iPSC-derived cardiomyocytes and neurons, dynamic characterization of iPSC-derived 3D cerebral organoids, CRISPR/Cas9 genome editing, and non-viral minicircle vector-based gene modification of stem cells. Different applications of stem cells in disease modeling are described as well. This volume also highlights the most recent developments and applications of stem cells in basic science research and disease treatments.
- Published
- 2020
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13. In vitro modeling for inherited neurological diseases using induced pluripotent stem cells: from 2D to organoid.
- Author
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Nam KH, Yi SA, Jang HJ, Han JW, and Lee J
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- Animals, Brain pathology, Cell Differentiation physiology, Disease Models, Animal, Humans, Induced Pluripotent Stem Cells physiology, Malformations of Cortical Development, Group I genetics, Mice, Neurodevelopmental Disorders genetics, Neurogenesis genetics, Neurons pathology, Neurons transplantation, Organoids pathology, Organoids physiology, Transplantation Chimera, Brain cytology, Cell Culture Techniques methods, Malformations of Cortical Development, Group I pathology, Neurodevelopmental Disorders pathology, Neurons physiology
- Abstract
Stem cells are characterized by self-renewal and by their ability to differentiate into cells of various organs. With massive progress in 2D and 3D cell culture techniques, in vitro generation of various types of such organoids from patient-derived stem cells is now possible. As in vitro differentiation protocols are usually made to resemble human developmental processes, organogenesis of patient-derived stem cells can provide key information regarding a range of developmental diseases. Human stem cell-based in vitro modeling as opposed to using animal models can particularly benefit the evaluation of neurological diseases because of significant differences in structure and developmental processes between the human and the animal brain. This review focuses on stem cell-based in vitro modeling of neurodevelopmental disorders, more specifically, the fundamentals and technical advancements in monolayer neuron and brain organoid cultures. Furthermore, we discuss the drawbacks of the conventional culture method and explore the advanced, cutting edge 3D organoid models for several neurodevelopmental diseases, including genetic diseases such as Down syndrome, Rett syndrome, and Miller-Dieker syndrome, as well as brain malformations like macrocephaly and microcephaly. Finally, we discuss the limitations of the current organoid techniques and some potential solutions that pave the way for accurate modeling of neurological disorders in a dish.
- Published
- 2020
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14. Optogenetics for neural transplant manipulation and functional analysis.
- Author
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Habibey R, Sharma K, Swiersy A, and Busskamp V
- Subjects
- Animals, Brain cytology, Brain physiology, Humans, Neural Stem Cells cytology, Neural Stem Cells metabolism, Neurons cytology, Neurons metabolism, Spinal Cord cytology, Spinal Cord physiology, Stem Cell Transplantation methods, Neural Stem Cells transplantation, Neurons transplantation, Optogenetics methods
- Abstract
Transplantation of neural stem cells (NSCs) or NSC-derived neurons into the brain is a promising therapeutic approach to restore neuronal function. Rapid progress in the NSCs research field, particularly due to the exploitation of induced pluripotent stem cells (iPSCs), offers great potential and an unlimited source of stem cell-derived neural grafts. Studying the functional integration of these grafts into host brain tissues and their effects on each other have been boosted by the implementation of optogenetic technologies. Optogenetics provides high spatiotemporal functional manipulations of grafted or host neurons in parallel. This review aims to highlight the impact of optogenetics in neural stem cell transplantations., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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15. Human Pluripotent Stem Cell-Derived Neurons Are Functionally Mature In Vitro and Integrate into the Mouse Striatum Following Transplantation.
- Author
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Comella-Bolla A, Orlandi JG, Miguez A, Straccia M, García-Bravo M, Bombau G, Galofré M, Sanders P, Carrere J, Segovia JC, Blasi J, Allen ND, Alberch J, Soriano J, and Canals JM
- Subjects
- Animals, Cell Line, Corpus Striatum cytology, Humans, Mice, Cell Culture Techniques methods, Corpus Striatum surgery, Neurogenesis physiology, Neurons transplantation, Pluripotent Stem Cells cytology
- Abstract
Human pluripotent stem cells (hPSCs) are a powerful tool for modelling human development. In recent years, hPSCs have become central in cell-based therapies for neurodegenerative diseases given their potential to replace affected neurons. However, directing hPSCs into specific neuronal types is complex and requires an accurate protocol that mimics endogenous neuronal development. Here we describe step-by-step a fast feeder-free neuronal differentiation protocol to direct hPSCs to mature forebrain neurons in 37 days in vitro (DIV). The protocol is based upon a combination of specific morphogens, trophic and growth factors, ions, neurotransmitters and extracellular matrix elements. A human-induced PSC line (Ctr-Q33) and a human embryonic stem cell line (GEN-Q18) were used to reinforce the potential of the protocol. Neuronal activity was analysed by single-cell calcium imaging. At 8 DIV, we obtained a homogeneous population of hPSC-derived neuroectodermal progenitors which self-arranged in bi-dimensional neural tube-like structures. At 16 DIV, we generated hPSC-derived neural progenitor cells (NPCs) with mostly a subpallial identity along with a subpopulation of pallial NPCs. Terminal in vitro neuronal differentiation was confirmed by the expression of microtubule associated protein 2b (Map 2b) by almost 100% of hPSC-derived neurons and the expression of specific-striatal neuronal markers including GABA, CTIP2 and DARPP-32. HPSC-derived neurons showed mature and functional phenotypes as they expressed synaptic markers, voltage-gated ion channels and neurotransmitter receptors. Neurons displayed diverse spontaneous activity patterns that were classified into three major groups, namely "high", "intermediate" and "low" firing neurons. Finally, transplantation experiments showed that the NPCs survived and differentiated within mouse striatum for at least 3 months. NPCs integrated host environmental cues and differentiated into striatal medium-sized spiny neurons (MSNs), which successfully integrated into the endogenous circuitry without teratoma formation. Altogether, these findings demonstrate the potential of this robust human neuronal differentiation protocol, which will bring new opportunities for the study of human neurodevelopment and neurodegeneration, and will open new avenues in cell-based therapies, pharmacological studies and alternative in vitro toxicology.
- Published
- 2020
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16. Injured adult neurons regress to an embryonic transcriptional growth state.
- Author
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Poplawski GHD, Kawaguchi R, Van Niekerk E, Lu P, Mehta N, Canete P, Lie R, Dragatsis I, Meves JM, Zheng B, Coppola G, and Tuszynski MH
- Subjects
- Animals, Axons pathology, Axons physiology, Disease Models, Animal, Female, Gene Expression Profiling, Huntingtin Protein genetics, Mice, Neural Stem Cells transplantation, Neuronal Plasticity, Neurons cytology, Neurons transplantation, Protein Biosynthesis, Pyramidal Tracts cytology, Pyramidal Tracts metabolism, Pyramidal Tracts pathology, RNA-Seq, Spinal Cord Injuries genetics, Spinal Cord Injuries pathology, Transcriptome, Cell Proliferation genetics, Embryo, Mammalian cytology, Embryo, Mammalian metabolism, Nerve Regeneration genetics, Neural Stem Cells cytology, Neurons metabolism, Neurons pathology, Transcription, Genetic
- Abstract
Grafts of spinal-cord-derived neural progenitor cells (NPCs) enable the robust regeneration of corticospinal axons and restore forelimb function after spinal cord injury
1 ; however, the molecular mechanisms that underlie this regeneration are unknown. Here we perform translational profiling specifically of corticospinal tract (CST) motor neurons in mice, to identify their 'regenerative transcriptome' after spinal cord injury and NPC grafting. Notably, both injury alone and injury combined with NPC grafts elicit virtually identical early transcriptomic responses in host CST neurons. However, in mice with injury alone this regenerative transcriptome is downregulated after two weeks, whereas in NPC-grafted mice this transcriptome is sustained. The regenerative transcriptome represents a reversion to an embryonic transcriptional state of the CST neuron. The huntingtin gene (Htt) is a central hub in the regeneration transcriptome; deletion of Htt significantly attenuates regeneration, which shows that Htt has a key role in neural plasticity after injury.- Published
- 2020
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17. Activity in grafted human iPS cell-derived cortical neurons integrated in stroke-injured rat brain regulates motor behavior.
- Author
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Palma-Tortosa S, Tornero D, Grønning Hansen M, Monni E, Hajy M, Kartsivadze S, Aktay S, Tsupykov O, Parmar M, Deisseroth K, Skibo G, Lindvall O, and Kokaia Z
- Subjects
- Action Potentials physiology, Animals, Behavior Observation Techniques, Behavior, Animal physiology, Cell Differentiation physiology, Cell Line, Disease Models, Animal, Humans, Infarction, Middle Cerebral Artery etiology, Infarction, Middle Cerebral Artery pathology, Infarction, Middle Cerebral Artery physiopathology, Male, Neurons physiology, Optogenetics, Patch-Clamp Techniques, Rats, Recovery of Function, Somatosensory Cortex cytology, Somatosensory Cortex pathology, Induced Pluripotent Stem Cells physiology, Infarction, Middle Cerebral Artery therapy, Motor Activity physiology, Neurons transplantation, Somatosensory Cortex physiopathology
- Abstract
Stem cell transplantation can improve behavioral recovery after stroke in animal models but whether stem cell-derived neurons become functionally integrated into stroke-injured brain circuitry is poorly understood. Here we show that intracortically grafted human induced pluripotent stem (iPS) cell-derived cortical neurons send widespread axonal projections to both hemispheres of rats with ischemic lesions in the cerebral cortex. Using rabies virus-based transsynaptic tracing, we find that at 6 mo after transplantation, host neurons in the contralateral somatosensory cortex receive monosynaptic inputs from grafted neurons. Immunoelectron microscopy demonstrates myelination of the graft-derived axons in the corpus callosum and that their terminals form excitatory, glutamatergic synapses on host cortical neurons. We show that the stroke-induced asymmetry in a sensorimotor (cylinder) test is reversed by transplantation. Light-induced inhibition of halorhodopsin-expressing, grafted neurons does not recreate the impairment, indicating that its reversal is not due to neuronal activity in the graft. However, we find bilateral decrease of motor performance in the cylinder test after light-induced inhibition of either grafted or endogenous halorhodopsin-expressing cortical neurons, located in the same area, and after inhibition of endogenous halorhodopsin-expressing cortical neurons by exposure of their axons to light on the contralateral side. Our data indicate that activity in the grafted neurons, probably mediated through transcallosal connections to the contralateral hemisphere, is involved in maintaining normal motor function. This is an example of functional integration of efferent projections from grafted neurons into the stroke-affected brain's neural circuitry, which raises the possibility that such repair might be achievable also in humans affected by stroke., Competing Interests: The authors declare no competing interest.
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- 2020
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18. 3D Bioprinting Electrically Conductive Bioink with Human Neural Stem Cells for Human Neural Tissues.
- Author
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Tomaskovic-Crook E and Crook JM
- Subjects
- Calcium analysis, Cells, Cultured, Electric Conductivity, Electric Stimulation, Fluorescent Dyes, Humans, Immunophenotyping, Microscopy, Confocal methods, Neurogenesis, Neuroglia transplantation, Neurons transplantation, Single-Cell Analysis, Biocompatible Materials, Bioprinting, Neural Stem Cells transplantation, Printing, Three-Dimensional, Tissue Engineering methods
- Abstract
Bioprinting cells with an electrically conductive bioink provides an opportunity to produce three-dimensional (3D) cell-laden constructs with the option of electrically stimulating cells in situ during and after tissue development. We and others have demonstrated the use of electrical stimulation (ES) to influence cell behavior and function for a more biomimetic approach to tissue engineering. Here, we detail a previously published method for 3D printing an electrically conductive bioink with human neural stem cells (hNSCs) that are subsequently differentiated. The differentiated tissue constructs comprise functional neurons and supporting neuroglia and are amenable to ES for the purposeful modulation of neural activity. Importantly, the method could be adapted to fabricate and stimulate neural and nonneural tissues from other cell types, with the potential to be applied for both research- and clinical-product development.
- Published
- 2020
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19. Transplantation of neural scaffolds consisting of dermal fibroblast-reprogrammed neurons and 3D silk fibrous materials promotes the repair of spinal cord injury.
- Author
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Hu Y, Zhang F, Zhong W, Liu Y, He Q, Yang M, Chen H, Xu X, Bian K, Xu J, Li J, Shen Y, and Zhang H
- Subjects
- Animals, Axons physiology, Biocompatible Materials chemistry, Biocompatible Materials pharmacology, Blood-Brain Barrier metabolism, Cell Transdifferentiation drug effects, Cells, Cultured, Cellular Reprogramming, Female, Fibroblasts cytology, Humans, Mice, Neurons cytology, Neurons metabolism, Porosity, Rats, Rats, Sprague-Dawley, Regeneration physiology, Nanofibers chemistry, Neurons transplantation, Silk chemistry, Spinal Cord Injuries therapy, Tissue Scaffolds chemistry
- Abstract
Transplantation of tissue-engineered neural scaffolds bears great potential for reconstructing neural circuits after spinal cord injury (SCI). In this study, a 3D porous silk fibrous scaffold (3D-SF) with biomimetic interconnected micro- to nanofibrous structure and good biocompatibility is fabricated. Then, a small-molecule combination CFLSSVY (CHIR99021, Forskolin, LDN193189, SB431542, SP600125, VPA, and Y27632) that efficiently reprograms rat dermal fibroblasts into neurons is screened, and these chemically induced neurons (CiNs) are shown to readily communicate on the 3D-SF and form neural scaffolds. After transplantation of these silk-based neural scaffolds into the stumps of transected spinal cords in rats, the damaged tissue is repaired significantly, as indicated by the reduced cavity areas, decreased GFAP expression, and improved axonal regeneration and myelination in the injury site. Moreover, the hindlimb movement and motor-nerve conductivity are greatly improved as indicated by the elevated BBB score, the alternate movement of two hindlimbs during the 45° inclined grid test, and the shortened latency and enhanced amplitude in cMEP detection. Together, these results demonstrate that transplantation of neural scaffolds consisting of 3D-SF and dermal fibroblast-reprogrammed neurons leads to significant nerve regeneration and functional recovery, providing a promising therapeutic strategy for SCI.
- Published
- 2019
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20. Enteric neuronal cell therapy reverses architectural changes in a novel diphtheria toxin-mediated model of colonic aganglionosis.
- Author
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Bhave S, Arciero E, Baker C, Ho WL, Stavely R, Goldstein AM, and Hotta R
- Subjects
- Animals, Colon cytology, Colon pathology, Diphtheria Toxin metabolism, Diphtheria Toxin toxicity, Disease Models, Animal, Heparin-binding EGF-like Growth Factor genetics, Heparin-binding EGF-like Growth Factor metabolism, Hirschsprung Disease genetics, Hirschsprung Disease pathology, Humans, Intestinal Mucosa cytology, Intestinal Mucosa pathology, Mice, Mice, Transgenic, Neural Crest cytology, Enteric Nervous System cytology, Hirschsprung Disease therapy, Neurons transplantation, Stem Cell Transplantation methods
- Abstract
Hirschsprung disease (HSCR) is characterized by absence of the enteric nervous system (ENS) in the distal bowel. Despite removal of the aganglionic segment, gastrointestinal (GI) problems persist. Cell therapy offers potential treatment but use of genetic models is limited by their poor survival. We have developed a novel model of aganglionosis in which enteric neural crest-derived cells (ENCDCs) express diphtheria toxin (DT) receptor. Local DT injection into the colon wall results in focal, specific, and sustained ENS ablation without altering GI transit or colonic contractility, allowing improved survival over other aganglionosis models. Focal ENS ablation leads to increased smooth muscle and mucosal thickness, and localized inflammation. Transplantation of ENCDCs into this region leads to engraftment, migration, and differentiation of enteric neurons and glial cells, with restoration of normal architecture of the colonic epithelium and muscle, reduction in inflammation, and improved survival.
- Published
- 2019
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21. The Combination of Adipose-derived Schwann-like Cells and Acellular Nerve Allografts Promotes Sciatic Nerve Regeneration and Repair through the JAK2/STAT3 Signaling Pathway in Rats.
- Author
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Fu XM, Wang Y, Fu WL, Liu DH, Zhang CY, Wang QL, and Tong XJ
- Subjects
- Animals, Brain-Derived Neurotrophic Factor biosynthesis, Ciliary Neurotrophic Factor biosynthesis, Male, Mesenchymal Stem Cell Transplantation methods, Nerve Growth Factor biosynthesis, Neurons transplantation, Rats, Recovery of Function physiology, Sciatic Nerve injuries, Sciatic Nerve surgery, Signal Transduction physiology, Spinal Cord metabolism, Allografts transplantation, Janus Kinase 2 physiology, Nerve Regeneration physiology, STAT3 Transcription Factor physiology, Sciatic Nerve physiopathology
- Abstract
Schwann cells (SCs) combined with acellular nerve allografts (ANAs) effectively promote the regeneration and repair of peripheral nerves, but the exact mechanism has not been fully elucidated. However, the disadvantages of SCs include their limited source and slow rate of expansion in vitro. Previous studies have found that adipose-derived stem cells have the ability to differentiate into Schwann-like cells. Therefore, we speculated that Schwann-like cells combined with ANAs could profoundly facilitate nerve regeneration and repair. The aim of the present study was to investigate the cellular and molecular mechanisms of regeneration and repair. In this study, tissue-engineered nerves were first constructed by adipose-derived Schwann-like cells and ANAs to bridge missing sciatic nerves. Then, the rats were randomly divided into five groups (n = 12 per group): a Control group; a Model group; an ADSC group; an SC-L group; and a DMEM group. Twelve weeks postsurgery, behavioral function tests and molecular biological techniques were used to evaluate the function of regenerated nerves and the relevant molecular mechanisms after sciatic nerve injury (SNI). The results showed that adipose-derived Schwann-like cells combined with ANAs markedly promoted sciatic nerve regeneration and repair. These findings also demonstrated that the expression of neurotrophic factors (NFs) was increased, and the expression of Janus activated kinase2 (JAK2)/P-JAK2, signal transducer and activator of transcription-3 (STAT3)/P-STAT3 was decreased in the spinal cord after SNI. Therefore, these results suggested that highly expressed NFs in the spinal cord could promote nerve regeneration and repair by inhibiting activation of the JAK2/STAT3 signaling pathway., (Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2019
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22. Rapid and Efficient Conversion of Human Fibroblasts into Functional Neurons by Small Molecules.
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Yang Y, Chen R, Wu X, Zhao Y, Fan Y, Xiao Z, Han J, Sun L, Wang X, and Dai J
- Subjects
- Animals, Cell Line, Fibroblasts drug effects, Humans, Mice, SCID, Neurons transplantation, Small Molecule Libraries pharmacology, Cellular Reprogramming drug effects, Cellular Reprogramming Techniques methods, Fibroblasts cytology, Neurogenesis drug effects, Neurons cytology
- Abstract
Recent studies have demonstrated that human astrocytes and fibroblasts can be directly converted into functional neurons by small molecules. However, fibroblasts, as a potentially better cell resource for transplantation, are not as easy to reprogram as astrocytes regarding their fate to neurons, and chemically induced neurons (iNs) with low efficiency from fibroblasts resulted in limited application for the treatment of neurological disorders, including depression. Here, we report that human fibroblasts can be efficiently and directly reprogrammed into glutamatergic neuron-like cells by serially exposing cells to a combination of small molecules. These iNs displayed neuronal transcriptional networks, and also exhibited mature firing patterns and formed functional synapses. Importantly, iNs could integrate into local circuits after transplantation into postnatal mouse brain. Our study provides a rapid and efficient transgene-free approach for chemically generating neuron-like cells from human fibroblasts. Furthermore, our approach offers strategies for disease modeling and drug discovery in central nervous system disorders., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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23. Plasmonic nano surface for neuronal differentiation and manipulation.
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Pandanaboina SC, Alghazali KM, Nima ZA, Alawajji RA, Sharma KD, Watanabe F, Saini V, Biris AS, and Srivatsan M
- Subjects
- Animals, Astrocytes transplantation, Brain Injuries, Traumatic pathology, Brain Injuries, Traumatic therapy, Cells, Cultured, Embryonic Stem Cells drug effects, Gold chemistry, Gold pharmacology, Humans, Neural Stem Cells drug effects, Neural Stem Cells transplantation, Neurodegenerative Diseases pathology, Neurons drug effects, Rats, Cell Differentiation drug effects, Nanotubes chemistry, Neurodegenerative Diseases therapy, Neurons transplantation
- Abstract
Neurodegenerative diseases and traumatic brain injuries can destroy neurons, resulting in sensory and motor function loss. Transplantation of differentiated neurons from stem cells could help restore such lost functions. Plasmonic gold nanorods (AuNR) were integrated in growth surfaces to stimulate and modulate neural cells in order to tune cell physiology. An AuNR nanocomposite system was fabricated, characterized, and then utilized to study the differentiation of embryonic rat neural stem cells (NSCs). Results demonstrated that this plasmonic surface 1) accelerated differentiation, yielding almost twice as many differentiated neural cells as a traditional NSC culture surface coated with poly-D-lysine and laminin for the same time period; and 2) promoted differentiation of NSCs into neurons and astrocytes in a 2:1 ratio, as evidenced by the expression of relevant marker proteins. These results indicate that the design and properties of this AuNR plasmonic surface would be advantageous for tissue engineering to address neural degeneration., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
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24. MHC matching fails to prevent long-term rejection of iPSC-derived neurons in non-human primates.
- Author
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Aron Badin R, Bugi A, Williams S, Vadori M, Michael M, Jan C, Nassi A, Lecourtois S, Blancher A, Cozzi E, Hantraye P, and Perrier AL
- Subjects
- Animals, Cell Differentiation immunology, Cytotoxicity, Immunologic immunology, Disease Models, Animal, Histocompatibility Testing, Humans, Huntington Disease immunology, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells immunology, Neurons cytology, Neurons immunology, Primates, Rats, Nude, Transplantation, Autologous, Graft Rejection immunology, Huntington Disease therapy, Induced Pluripotent Stem Cells transplantation, Major Histocompatibility Complex immunology, Neurons transplantation
- Abstract
Cell therapy products (CTP) derived from pluripotent stem cells (iPSCs) may constitute a renewable, specifically differentiated source of cells to potentially cure patients with neurodegenerative disorders. However, the immunogenicity of CTP remains a major issue for therapeutic approaches based on transplantation of non-autologous stem cell-derived neural grafts. Despite its considerable side-effects, long-term immunosuppression, appears indispensable to mitigate neuro-inflammation and prevent rejection of allogeneic CTP. Matching iPSC donors' and patients' HLA haplotypes has been proposed as a way to access CTP with enhanced immunological compatibility, ultimately reducing the need for immunosuppression. In the present work, we challenge this paradigm by grafting autologous, MHC-matched and mis-matched neuronal grafts in a primate model of Huntington's disease. Unlike previous reports in unlesioned hosts, we show that in the absence of immunosuppression MHC matching alone is insufficient to grant long-term survival of neuronal grafts in the lesioned brain.
- Published
- 2019
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25. Controlled Cortical Impact Model of Mouse Brain Injury with Therapeutic Transplantation of Human Induced Pluripotent Stem Cell-Derived Neural Cells.
- Author
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Furmanski O, Nieves MD, and Doughty ML
- Subjects
- Animals, Behavior, Animal, Brain Injuries pathology, Brain Injuries, Traumatic pathology, Cells, Cultured, Craniotomy, Disease Models, Animal, Humans, Male, Mice, Monitoring, Intraoperative, Brain Injuries therapy, Cerebral Cortex pathology, Induced Pluripotent Stem Cells cytology, Neurons transplantation
- Abstract
Traumatic brain injury (TBI) is a leading cause of morbidity and mortality worldwide. Disease pathology due to TBI progresses from the primary mechanical insult to secondary injury processes, including apoptosis and inflammation. Animal modeling has been valuable in the search to unravel injury mechanisms and evaluate potential neuroprotective therapies. This protocol describes the controlled cortical impact (CCI) model of focal, open-head TBI. Specifically, parameters for producing a mild unilateral cortical injury are described. Behavioral consequences of CCI are analyzed using the adhesive tape removal test of bilateral sensorimotor integration. Regarding experimental therapy for TBI pathology, this protocol also illustrates a process for transplanting cultured cells into the brain. Neural cell cultures derived from human induced pluripotent stem cells (hiPSCs) were chosen for their potential to show superior functional restoration in human TBI patients. Chronic survival of hiPSCs in the host mouse brain tissue is detected using a modified DAB immunohistochemical process.
- Published
- 2019
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26. Primary tissue for cellular brain repair in Parkinson's disease: Promise, problems and the potential of biomaterials.
- Author
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Moriarty N, Parish CL, and Dowd E
- Subjects
- Animals, Humans, Biocompatible Materials therapeutic use, Fetal Tissue Transplantation, Neurons transplantation, Parkinson Disease therapy
- Abstract
The dopamine precursor, levodopa, remains the "gold standard" treatment for Parkinson's disease, and, although it provides superlative efficacy in the early stages of the disease, its long-term use is limited by the development of severe motor side effects and a significant abating of therapeutic efficacy. Therefore, there remains a major unmet clinical need for the development of effective neuroprotective, neurorestorative or neuroreparatory therapies for this condition. The relatively selective loss of dopaminergic neurons from the nigrostriatal pathway makes Parkinson's disease an ideal candidate for reparative cell therapies, wherein the dopaminergic neurons that are lost in the condition are replaced through direct cell transplantation into the brain. To date, this approach has been developed, validated and clinically assessed using dopamine neuron-rich foetal ventral mesencephalon grafts which have been shown to survive and reinnervate the denervated brain after transplantation, and to restore motor function. However, despite long-term symptomatic relief in some patients, significant limitations, including poor graft survival and the impact this has on the number of foetal donors required, have prevented this therapy being more widely adopted as a restorative approach for Parkinson's disease. Injectable biomaterial scaffolds have the potential to improve the delivery, engraftment and survival of these grafts in the brain through provision of a supportive microenvironment for cell adhesion, growth and immune shielding. This article will briefly review the development of primary cell therapies for brain repair in Parkinson's disease and will consider the emerging literature which highlights the potential of using injectable biomaterial hydrogels in this context., (© 2018 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)
- Published
- 2019
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27. High efficient differentiation of human adipose-derived stem cells into retinal pigment epithelium-like cells in medium containing small molecules inducers with a simple method.
- Author
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Aboutaleb Kadkhodaeian H, Salati A, and Lashay A
- Subjects
- Cell Shape, Cells, Cultured, Culture Media chemistry, Humans, Mesenchymal Stem Cells drug effects, Neurons drug effects, Neurons transplantation, Retina drug effects, Retina pathology, Retina transplantation, Retinal Degeneration pathology, Retinal Pigment Epithelium cytology, Small Molecule Libraries administration & dosage, Cell Differentiation drug effects, Mesenchymal Stem Cell Transplantation, Mesenchymal Stem Cells cytology, Retinal Degeneration therapy, Retinal Pigment Epithelium transplantation
- Abstract
Background: The induction of retinal pigmented epithelium cells (RPE) is one of the most important objectives in research focused on treating retinal degenerative diseases. The present study aims to differentiate human adipose stem cells (hADSCs) into RPE cells for replacement therapies in cases of retinal degenerative diseases., Methods: Lipoaspirate-derived human adipose stem cells (LA-hADSCs) were obtained from abdominal samples and examined by immunocytochemistry for the expression of mesenchymal adipose stem cell markers. RPE cells were also obtained from human samples and cultured to be used as control after being examined for the expression of their designated markers. hADSCs differentiated into RPE cells after 80 days using chemical inducers in one steps. The differentiated cells were then compared to control cells in marker expression. The differentiated cells were also examined under a scanning electron microscope for the presence of apical microvilli and cell connection., Results: Cultured hADSCs at the fourth passage was shown to express the surface markers CD90 (98 ± 2%), CD11b (96 ± 3%), and CD105 (95 ± 4%). The RPE cells obtained from human samples expressed the marker RPE65 quite well. 80 days after differentiation, the previously hADSCs expressed both RPE65 (100%) and CRALBP (96 ± 1%) and were thus significantly similar to the RPE cells obtained from human samples. Morphologically, differentiated cells appeared to have epithelial and cytoplasmic pigment granules. Observations using a scanning electron microscope recorded clear connections among the differentiated RPE cells and revealed apical microvilli., Conclusion: Human adipose stem cells can differentiate into retinal pigmented epithelium cells, which can be used in cell replacement therapy for degenerative diseases including age-related macular degeneration (AMD) as well as retinitis pigmentosa (RP)., (Published by Elsevier Ltd.)
- Published
- 2019
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28. Induced Neurons for the Study of Neurodegenerative and Neurodevelopmental Disorders.
- Author
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Sauter EJ, Kutsche LK, Klapper SD, and Busskamp V
- Subjects
- Cell Differentiation, Coculture Techniques, Humans, Induced Pluripotent Stem Cells transplantation, Neurodegenerative Diseases genetics, Neurodevelopmental Disorders genetics, Neurons transplantation, Transcription Factors antagonists & inhibitors, Astrocytes cytology, CRISPR-Cas Systems, Induced Pluripotent Stem Cells cytology, Neurodegenerative Diseases therapy, Neurodevelopmental Disorders therapy, Neurons cytology, Transcription Factors genetics
- Abstract
Patient-derived or genomically modified human induced pluripotent stem cells (iPSCs) offer the opportunity to study neurodevelopmental and neurodegenerative disorders. Overexpression of certain neurogenic transcription factors (TFs) in iPSCs can induce efficient differentiation into homogeneous populations of the disease-relevant neuronal cell types. Here we provide protocols for genomic manipulations of iPSCs by CRISPR/Cas9. We also introduce two methods, based on lentiviral delivery and the piggyBac transposon system, to stably integrate neurogenic TFs into human iPSCs. Furthermore, we describe the TF-mediated neuronal differentiation and maturation in combination with astrocyte cocultures.
- Published
- 2019
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29. Minimally Invasive and Regenerative Therapeutics.
- Author
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Ashammakhi N, Ahadian S, Darabi MA, El Tahchi M, Lee J, Suthiwanich K, Sheikhi A, Dokmeci MR, Oklu R, and Khademhosseini A
- Subjects
- Biocompatible Materials chemistry, Biocompatible Materials therapeutic use, Humans, Nanoparticles chemistry, Neurons cytology, Neurons transplantation, Robotics, Spinal Cord cytology, Spinal Cord transplantation, Stem Cell Transplantation, Stem Cells cytology, Stem Cells metabolism, Tissue Engineering, Regenerative Medicine
- Abstract
Advances in biomaterial synthesis and fabrication, stem cell biology, bioimaging, microsurgery procedures, and microscale technologies have made minimally invasive therapeutics a viable tool in regenerative medicine. Therapeutics, herein defined as cells, biomaterials, biomolecules, and their combinations, can be delivered in a minimally invasive way to regenerate different tissues in the body, such as bone, cartilage, pancreas, cardiac, skeletal muscle, liver, skin, and neural tissues. Sophisticated methods of tracking, sensing, and stimulation of therapeutics in vivo using nano-biomaterials and soft bioelectronic devices provide great opportunities to further develop minimally invasive and regenerative therapeutics (MIRET). In general, minimally invasive delivery methods offer high yield with low risk of complications and reduced costs compared to conventional delivery methods. Here, minimally invasive approaches for delivering regenerative therapeutics into the body are reviewed. The use of MIRET to treat different tissues and organs is described. Although some clinical trials have been performed using MIRET, it is hoped that such therapeutics find wider applications to treat patients. Finally, some future perspective and challenges for this emerging field are highlighted., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2019
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30. Preconditioned neurons with NaB and nicorandil, a favorable source for stroke cell therapy.
- Author
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Hosseini SM, Ziaee SM, Haider KH, Karimi A, Tabeshmehr P, and Abbasi Z
- Subjects
- Animals, Brain-Derived Neurotrophic Factor genetics, Butyric Acid pharmacology, Cell Survival drug effects, Cell- and Tissue-Based Therapy methods, Cellular Microenvironment genetics, DNA-(Apurinic or Apyrimidinic Site) Lyase genetics, Humans, Mice, NF-kappa B genetics, Neural Stem Cells drug effects, Neurons pathology, Nicorandil pharmacology, Phosphatidylinositol 3-Kinases genetics, Stroke genetics, Stroke pathology, Cellular Microenvironment drug effects, Neural Stem Cells transplantation, Neurons transplantation, Stroke therapy
- Abstract
Poor survival of stem cells in the harsh microenvironment at the site of stroke, especially during acute phase of injury, remains a serious obstacle to achieve the desired prognosis. We hypothesized that combined treatment of neural stem cells (NSCs) with small molecules would precondition them to become robust and survive better as compared with the native nonpreconditioned cells. Mouse ganglionic NSCs were isolated, cultured, and characterized. The cells were preconditioned by treatment with sodium butyrate (NaB) and nicorandil (Nico) and transplanted in an experimentally induced stroke model. Sham-operated animals without treatment or animals with experimental stroke treated with basal medium, native NSCs, NSCs preconditioned with NaB or Nico alone were used as controls. The tissue samples and cells with different treatments were used to measure brain-tissue-derived neurotrophic factor (BDNF) level and the activity of phosphatidylinositol-3 kinase (PI3K), apurinic/apyrimidinic endonuclease 1 (APE1), and nuclear factor-κB (NF-κB) p50 both in vitro and in vivo, respectively. Additionally, survival of the cells and recovery indices for stroke were studied. The combined treatment with NaB + Nico resulted in increased BDNF level and higher PI3K, APE1, and the downstream NF-κB activation, which were blocked by pretreatment with their respective inhibitors. Donor cell survival increased postengraftment as assessed by 5-bromo-2'-deoxyuridine immunostaining and reduced Terminal deoxynucleotide transferase dUTP Nick End Labeling positivity at the site of engraftment. There was reduction in proinflammatory cytokines and infiltration of both GFAP
+ and CD68+ at the injury site. There was reduction in the infarct size and neurological function was preserved in the preconditioned cell treatment group. Our preconditioning approach with small molecules effectively improved the survival as well as functionality of NSCs., (© 2018 Wiley Periodicals, Inc.)- Published
- 2018
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31. Induction of human somatostatin and parvalbumin neurons by expressing a single transcription factor LIM homeobox 6.
- Author
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Yuan F, Chen X, Fang KH, Wang Y, Lin M, Xu SB, Huo HQ, Xu M, Ma L, Chen Y, He S, and Liu Y
- Subjects
- Action Potentials, Animals, Animals, Newborn, Body Patterning, Cell Differentiation, Cell Line, Gene Expression Profiling, Humans, Interneurons cytology, Interneurons metabolism, Mice, SCID, Neurons cytology, Neurons transplantation, Pluripotent Stem Cells cytology, Pluripotent Stem Cells metabolism, Prosencephalon cytology, gamma-Aminobutyric Acid metabolism, LIM-Homeodomain Proteins metabolism, Nerve Tissue Proteins metabolism, Neurons metabolism, Parvalbumins metabolism, Somatostatin metabolism, Transcription Factors metabolism
- Abstract
Human GABAergic interneurons (GIN) are implicated in normal brain function and in numerous mental disorders. However, the generation of functional human GIN subtypes from human pluripotent stem cells (hPSCs) has not been established. By expressing LHX6, a transcriptional factor that is critical for GIN development, we induced hPSCs to form GINs, including somatostatin (SST, 29%) and parvalbumin (PV, 21%) neurons. Our RNAseq results also confirmed the alteration of GIN identity with the overexpression of LHX6 . Five months after transplantation into the mouse brain, the human GABA precursors generated increased population of SST and PV neurons by overexpressing LHX6 . Importantly, the grafted human GINs exhibited functional electrophysiological properties and even fast-spiking-like action potentials. Thus, expression of the single transcription factor LHX6 under our GIN differentiation condition is sufficient to robustly induce human PV and SST subtypes., Competing Interests: FY, XC, KF, YW, ML, SX, HH, MX, LM, YC, SH, YL No competing interests declared, (© 2018, Yuan et al.)
- Published
- 2018
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32. A Chemical Recipe for Generation of Clinical-Grade Striatal Neurons from hESCs.
- Author
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Wu M, Zhang D, Bi C, Mi T, Zhu W, Xia L, Teng Z, Hu B, and Wu Y
- Subjects
- Animals, Cell Line, Coculture Techniques methods, Humans, Huntington Disease therapy, Mice, Mice, SCID, Neurons transplantation, Amyloid Precursor Protein Secretases antagonists & inhibitors, Corpus Striatum cytology, Diamines pharmacology, Human Embryonic Stem Cells cytology, Human Embryonic Stem Cells drug effects, Neurogenesis drug effects, Neurons cytology, Thiazoles pharmacology
- Abstract
Differentiation of human pluripotent stem cells (hPSCs) into striatal medium spiny neurons (MSNs) promises a cell-based therapy for Huntington's disease. However, clinical-grade MSNs remain unavailable. Here, we developed a chemical recipe named XLSBA to generate clinical-grade MSNs from embryonic stem cells (ESCs). We introduced the γ-secretase inhibitor DAPT into the recipe to accelerate neural differentiation, and replaced protein components with small molecules. Using this optimized protocol we could efficiently direct regular human ESCs (hESCs) as well as clinical-grade hESCs to lateral ganglionic eminence (LGE)-like progenitors and striatal MSNs within less than half of the time than previous protocols (within 14 days and 21 days, respectively). These striatal cells expressed appropriate MSN markers and electrophysiologically acted like authentic MSNs. Upon transplantation into brains of neonatal mice or mouse model of Huntington's disease, they exhibited sufficient safety and reasonable efficacy. Therefore, this quick and highly efficient derivation of MSNs offers unprecedented access to clinical application., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
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33. [Restoration of damaged cortical pathways by neural grafting].
- Author
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Ballout N, Péron S, and Gaillard A
- Subjects
- Adult, Animals, Brain Injuries physiopathology, Brain Injuries surgery, Humans, Motor Cortex injuries, Nerve Fibers transplantation, Motor Cortex pathology, Nerve Net surgery, Nerve Regeneration physiology, Neurons transplantation
- Abstract
The motor cortex plays a central role in the control, planning, and execution of voluntary motor commands in mammals. The loss of cortical neurons is a common feature of many neuropathological conditions such as traumatic and ischemic lesions or several neurodegenerative diseases. Cell transplantation presents a promising therapeutic strategy to overcome the limited abilities of axonal regrowth and spontaneous regeneration of the adult central nervous system. In this review, we will present a historical review of brain transplantation and the current state of research in the field of cortical transplantation., (© 2018 médecine/sciences – Inserm.)
- Published
- 2018
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34. Autologous Induced Pluripotent Stem Cell-Derived Neurons to Treat Parkinson's Disease.
- Author
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Loring JF
- Subjects
- Female, Humans, Induced Pluripotent Stem Cells cytology, Male, Neural Stem Cells transplantation, Parkinson Disease pathology, United States, United States Food and Drug Administration, Induced Pluripotent Stem Cells transplantation, Neurons transplantation, Parkinson Disease therapy, Transplantation, Autologous trends
- Abstract
In 2012, we planned a program to develop a neuron replacement therapy for Parkinson's disease (PD) that would have the greatest promise to help the patients. PD is a movement disorder caused by the progressive, inevitable loss of a specific type of dopamine neuron in the brain. The only viable treatment to reverse the progress of the disease is to replace those neurons; we decided to make dopamine neurons that matched the patients, by differentiating induced pluripotent stem cells that we generated from individuals with PD. This autologous cell therapy is entering the regulatory approval process this year with the U.S. Food and Drug Administration to begin to transplant the cells in the following 1 to 2 years.
- Published
- 2018
- Full Text
- View/download PDF
35. Three-dimensional brain-like microenvironments facilitate the direct reprogramming of fibroblasts into therapeutic neurons.
- Author
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Jin Y, Lee JS, Kim J, Min S, Wi S, Yu JH, Chang GE, Cho AN, Choi Y, Ahn DH, Cho SR, Cheong E, Kim YG, Kim HP, Kim Y, Kim DS, Kim HW, Quan Z, Kang HC, and Cho SW
- Subjects
- Animals, Cell Culture Techniques instrumentation, Cell Culture Techniques methods, Cell Transdifferentiation, Cellular Microenvironment, Disease Models, Animal, Fibroblasts cytology, Fibroblasts metabolism, Humans, Hydrogels chemistry, Locomotion, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Neovascularization, Physiologic, Neurons cytology, Neurons metabolism, Neurons transplantation, Recovery of Function, Stroke metabolism, Stroke pathology, Stroke therapy, Transcriptome, Brain metabolism, Cellular Reprogramming, Extracellular Matrix metabolism
- Abstract
Biophysical cues can improve the direct reprogramming of fibroblasts into neurons that can be used for therapeutic purposes. However, the effects of a three-dimensional (3D) environment on direct neuronal reprogramming remain unexplored. Here, we show that brain extracellular matrix (BEM) decellularized from human brain tissue facilitates the plasmid-transfection-based direct conversion of primary mouse embryonic fibroblasts into induced neuronal (iN) cells. We first show that two-dimensional (2D) surfaces modified with BEM significantly increase the generation efficiency of iN cells and enhance neuronal transdifferentiation and maturation. Moreover, in an animal model of ischaemic stroke, iN cells generated on the BEM substrates and transplanted into the brain led to significant improvements in locomotive behaviours. We also show that compared with the 2D BEM substrates, 3D BEM hydrogels recapitulating brain-like microenvironments further promote neuronal conversion and potentiate the functional recovery of the animals. Our findings suggest that 3D microenvironments can boost nonviral direct reprogramming for the generation of therapeutic neuronal cells.
- Published
- 2018
- Full Text
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36. Transplanted neural-like cells improve memory and Alzheimer-like pathology in a rat model.
- Author
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Hoveizi E, Mohammadi T, Moazedi AA, Zamani N, and Eskandary A
- Subjects
- Alzheimer Disease physiopathology, Animals, Basal Nucleus of Meynert, Cell Differentiation, Cell Survival, Disease Models, Animal, Embryoid Bodies cytology, Fibroblasts cytology, Fibroblasts metabolism, Male, Mice, Mouse Embryonic Stem Cells cytology, Mouse Embryonic Stem Cells metabolism, Nestin metabolism, Rats, Wistar, Alzheimer Disease pathology, Alzheimer Disease therapy, Memory, Neurons cytology, Neurons transplantation
- Abstract
Background Aims: Degeneration of the central nerve system, particularly in Alzheimer's disease, is a burden on society, and despite years of research, there is no effective treatment. Cell therapy appears to be an option that is of growing interest in neural studies. The main aim of this study was to investigate the histological and physiological effects of transplantation the neuron-like cell (NLC)-derived mouse embryonic stem cells (mESCs) on the repair of brain lesions in an Alzheimer's animal model (AM) in rats., Methods: Behavioral experiments were conducted in the light hours in a Y-shaped maze device. Animals were randomly divided into five groups, with seven rats per group. The nucleus basalis of Meynert (NBM) was destroyed bilaterally with an electrical lesion (0.5 mA for 3 s). One week after the bilateral lesion of the NBM, the differentiated NLCs (0.1 mL) were injected with stereotaxic surgery using a Hamilton syringe at NBM coordinates, and behavioral and histological tests were performed by the Y-maze task and hematoxylin and eosin staining after five weeks of the lesion. Also, differentiated cells detected by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis and fluorescent immunostaining., Results: The expression of neuronal markers including Nestin, Map2, NF-H, Tuj-1, GFAP and Olig-2 was surveyed by using the immunocytochemistry and qRT-PCR methods, and the results confirmed that the genes in question were expressed significantly more compared than the control sample. Five weeks after the cell transplantation in the AM, morphological and physiological investigation during the determination period confirmed improved disease state in the tested models., Conclusions: It should be noted that by improving the neuronal connectivity in AM rat brains, the transplanted NLCs rescue Alzheimer's cognition. This research has presented some preclinical evidence that showed NLCs transplantation can be used for AM treatment., (Copyright © 2018 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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37. Noninvasive imaging of nanoparticle-labeled transplant populations within polymer matrices for neural cell therapy.
- Author
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Tickle JA, Poptani H, Taylor A, and Chari DM
- Subjects
- Animals, Astrocytes chemistry, Astrocytes transplantation, Cell Proliferation drug effects, Cell Survival drug effects, Cell Tracking, Humans, Hydrogels chemistry, Hydrogels pharmacology, Magnetic Resonance Imaging, Magnetite Nanoparticles chemistry, Microscopy, Electron, Transmission, Nerve Degeneration drug therapy, Nerve Degeneration pathology, Neurons chemistry, Neurons transplantation, Polymers administration & dosage, Polymers chemistry, Rats, Astrocytes ultrastructure, Cell- and Tissue-Based Therapy, Magnetite Nanoparticles administration & dosage, Nerve Degeneration diagnostic imaging, Neurons ultrastructure
- Abstract
Aim: To develop a 3D neural cell construct for encapsulated delivery of transplant cells; develop hydrogels seeded with magnetic nanoparticle (MNP)-labeled cells suitable for cell tracking by MRI., Materials & Methods: Astrocytes were exogenously labeled with MRI-compatible iron-oxide MNPs prior to intra-construct incorporation within a 3D collagen hydrogel., Results: A connective, complex cellular network was clearly observable within the 3D constructs, with high cellular viability. MNP accumulation in astrocytes provided a hypointense MRI signal at 24 h & 14 days., Conclusion: Our findings support the concept of developing a 3D construct possessing the dual advantages of (i) support of long-term cell survival of neural populations with (ii) the potential for noninvasive MRI-tracking of intra-construct cells for neuroregenerative applications.
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- 2018
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38. Human Pluripotent Stem-Cell-Derived Cortical Neurons Integrate Functionally into the Lesioned Adult Murine Visual Cortex in an Area-Specific Way.
- Author
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Espuny-Camacho I, Michelsen KA, Linaro D, Bilheu A, Acosta-Verdugo S, Herpoel A, Giugliano M, Gaillard A, and Vanderhaeghen P
- Subjects
- Animals, Axons metabolism, Biomarkers metabolism, Cerebral Cortex cytology, Human Embryonic Stem Cells cytology, Humans, Mice, Inbred NOD, Mice, SCID, Organ Specificity, Synapses metabolism, Telencephalon metabolism, Aging pathology, Neurons transplantation, Pluripotent Stem Cells cytology, Visual Cortex pathology
- Abstract
The transplantation of pluripotent stem-cell-derived neurons constitutes a promising avenue for the treatment of several brain diseases. However, their potential for the repair of the cerebral cortex remains unclear, given its complexity and neuronal diversity. Here, we show that human visual cortical cells differentiated from embryonic stem cells can be transplanted and can integrate successfully into the lesioned mouse adult visual cortex. The transplanted human neurons expressed the appropriate repertoire of markers of six cortical layers, projected axons to specific visual cortical targets, and were synaptically active within the adult brain. Moreover, transplant maturation and integration were much less efficient following transplantation into the lesioned motor cortex, as previously observed for transplanted mouse cortical neurons. These data constitute an important milestone for the potential use of human PSC-derived cortical cells for the reassembly of cortical circuits and emphasize the importance of cortical areal identity for successful transplantation., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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39. Precocious deposition of perineuronal nets on Parvalbumin inhibitory neurons transplanted into adult visual cortex.
- Author
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Bradshaw KP, Figueroa Velez DX, Habeeb M, and Gandhi SP
- Subjects
- Age Factors, Animals, Cell Communication drug effects, Embryo, Mammalian, Female, Mice, Mice, Inbred C57BL, Mice, Transgenic, Nerve Net drug effects, Neurogenesis drug effects, Neurogenesis physiology, Neurons drug effects, Plant Lectins metabolism, Plant Lectins pharmacology, Pregnancy, Receptors, N-Acetylglucosamine metabolism, Time Factors, Visual Cortex drug effects, Cell Adhesion drug effects, Nerve Net metabolism, Neurons metabolism, Neurons transplantation, Parvalbumins metabolism, Visual Cortex cytology
- Abstract
The end of the critical period for primary visual cortex (V1) coincides with the deposition of perineuronal nets (PNN) onto Parvalbumin (PV) inhibitory neurons. Recently, we found that transplantation of embryonic inhibitory neurons into adult V1 reinstates a new critical period. Here we used Wisteria Floribunda Agglutinin (WFA) staining to compare the deposition of PNNs onto neurons during normal development and following transplantation at equivalent cell ages. In accord with previous findings, PV and PNN expression increases from negligible levels at postnatal day 14 (P14) to mature levels by P70. In contrast to P14, PNNs are found on transplanted PV neurons by 21 days after transplantation and persist to 105 days after transplantation. This precocious deposition was specific to PV neurons and excluded transplanted neurons expressing Somatostatin. Notably, the onset of PV expression in transplanted inhibitory neurons follows the timing of PV expression in juvenile V1. Moreover, transplantation has no discernible effect on host PNNs. The precocious deposition of PNNs onto transplanted PV neurons suggests that PNN expression identified by WFA does not reflect neuronal maturity and may be an inaccurate marker for transplant-induced plasticity of cortical circuits.
- Published
- 2018
- Full Text
- View/download PDF
40. Functional role of mesenchymal stem cells in the treatment of chronic neurodegenerative diseases.
- Author
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Lo Furno D, Mannino G, and Giuffrida R
- Subjects
- Alzheimer Disease physiopathology, Alzheimer Disease therapy, Amyotrophic Lateral Sclerosis physiopathology, Amyotrophic Lateral Sclerosis therapy, Humans, Huntington Disease pathology, Huntington Disease therapy, Mesenchymal Stem Cells cytology, Mesenchymal Stem Cells physiology, Neurodegenerative Diseases physiopathology, Neurons transplantation, Parkinson Disease physiopathology, Parkinson Disease therapy, Umbilical Cord transplantation, Cell- and Tissue-Based Therapy, Mesenchymal Stem Cell Transplantation, Neurodegenerative Diseases therapy, Neurogenesis physiology
- Abstract
Mesenchymal stem cells (MSCs) can differentiate into not only cells of mesodermal lineages, but also into endodermal and ectodermal derived elements, including neurons and glial cells. For this reason, MSCs have been extensively investigated to develop cell-based therapeutic strategies, especially in pathologies whose pharmacological treatments give poor results, if any. As in the case of irreversible neurological disorders characterized by progressive neuronal death, in which behavioral and cognitive functions of patients inexorably decline as the disease progresses. In this review, we focus on the possible functional role exerted by MSCs in the treatment of some disabling neurodegenerative disorders such as Alzheimer's Disease, Amyotrophic Lateral Sclerosis, Huntington's Disease, and Parkinson's Disease. Investigations have been mainly performed in vitro and in animal models by using MSCs generally originated from umbilical cord, bone marrow, or adipose tissue. Positive results obtained have prompted several clinical trials, the number of which is progressively increasing worldwide. To date, many of them have been primarily addressed to verify the safety of the procedures but some improvements have already been reported, fortunately. Although the exact mechanisms of MSC-induced beneficial activities are not entirely defined, they include neurogenesis and angiogenesis stimulation, antiapoptotic, immunomodulatory, and anti-inflammatory actions. Most effects would be exerted through their paracrine expression of neurotrophic factors and cytokines, mainly delivered at damaged regions, given the innate propensity of MSCs to home to injured sites. Hopefully, in the near future more efficacious cell-replacement therapies will be developed to substantially restore disease-disrupted brain circuitry., (© 2017 Wiley Periodicals, Inc.)
- Published
- 2018
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41. Injectable hydrogels of optimized acellular nerve for injection in the injured spinal cord.
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Cornelison RC, Gonzalez-Rothi EJ, Porvasnik SL, Wellman SM, Park JH, Fuller DD, and Schmidt CE
- Subjects
- Alginates chemistry, Animals, Axons physiology, Cell Movement, Glycosaminoglycans chemistry, Microscopy, Confocal, Rats, Schwann Cells, Spheroids, Cellular, Spinal Cord pathology, Tissue Engineering methods, Hydrogels chemistry, Nerve Regeneration, Neurons transplantation, Spinal Cord Injuries therapy
- Abstract
Objective: Spinal cord injury (SCI) affects a quarter million individuals in the United States, and there is currently no clinical treatment. Both fresh and acellular peripheral nerve grafts can induce spinal axon regeneration and support functional recovery in experimental injury models. Nonetheless, a scaffold that can be injected into a spinal contusion would be far less invasive to apply. We aimed to develop the first injectable acellular nerve graft for promoting repair after contusion SCI., Approach: We report a method to enzymatically solubilize optimized acellular (OA) nerve-a decellularized peripheral nerve graft developed in our laboratory and currently used clinically-to obtain an injectable solution that undergoes thermal gelation under physiological conditions. We quantified multiple physical and compositional properties of this novel material as well as tested its efficacy at acute and chronic time points following cervical contusion SCI., Main Results: This injectable optimized acellular (iOA) nerve graft retains native chemical cues such as collagens and glycosaminoglycans. By varying hydrogel concentration, the rheological properties and compressive modulus of iOA were similar to that previous reported for rat central nervous tissue. iOA solution was compatible with rat Schwann cells in culture, and hydrogel injection into a rat cervical contusion model significantly reduced the ratio of M1:M2 macrophages after one week, favoring regenerative phenotypes (p < 0.05). Furthermore, while iOA treatment did not affect locomotor or respiratory recovery over an eight week period, the percentage of axonal coverage increased at the distal tissue interface (p < 0.05), suggesting enhanced axonal extension within this region., Significance: Our data indicate that this novel injectable form of acellular nerve grafts is amenable for use after contusion SCI and may bolster a simultaneous therapy by acutely modulating the inflammatory milieu and supporting axonal growth.
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- 2018
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42. SOX10 Single Transcription Factor-Based Fast and Efficient Generation of Oligodendrocytes from Human Pluripotent Stem Cells.
- Author
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García-León JA, Kumar M, Boon R, Chau D, One J, Wolfs E, Eggermont K, Berckmans P, Gunhanlar N, de Vrij F, Lendemeijer B, Pavie B, Corthout N, Kushner SA, Dávila JC, Lambrichts I, Hu WS, and Verfaillie CM
- Subjects
- Amyotrophic Lateral Sclerosis genetics, Amyotrophic Lateral Sclerosis therapy, Antigens, Surface genetics, Gene Expression Regulation, Developmental, Humans, Multiple Sclerosis genetics, Multiple Sclerosis pathology, Multiple Sclerosis therapy, Myelin Basic Protein genetics, Neurons pathology, Neurons transplantation, Oligodendroglia cytology, Oligodendroglia transplantation, Pluripotent Stem Cells cytology, Pluripotent Stem Cells transplantation, Transcriptome genetics, Cell Differentiation genetics, Oligodendroglia metabolism, Pluripotent Stem Cells metabolism, SOXE Transcription Factors genetics
- Abstract
Scarce access to primary samples and lack of efficient protocols to generate oligodendrocytes (OLs) from human pluripotent stem cells (hPSCs) are hampering our understanding of OL biology and the development of novel therapies. Here, we demonstrate that overexpression of the transcription factor SOX10 is sufficient to generate surface antigen O4-positive (O4
+ ) and myelin basic protein-positive OLs from hPSCs in only 22 days, including from patients with multiple sclerosis or amyotrophic lateral sclerosis. The SOX10-induced O4+ population resembles primary human OLs at the transcriptome level and can myelinate neurons in vivo. Using in vitro OL-neuron co-cultures, myelination of neurons by OLs can also be demonstrated, which can be adapted to a high-throughput screening format to test the response of pro-myelinating drugs. In conclusion, we provide an approach to generate OLs in a very rapid and efficient manner, which can be used for disease modeling, drug discovery efforts, and potentially for therapeutic OL transplantation., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2018
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43. Nerve stepping stone has minimal impact in aiding regeneration across long acellular nerve allografts.
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Yan Y, Hunter DA, Schellhardt L, Ee X, Snyder-Warwick AK, Moore AM, Mackinnon SE, and Wood MD
- Subjects
- Aging, Allografts, Animals, Axons physiology, Biomarkers analysis, Gene Expression, Genetic Markers, Male, Muscle, Skeletal growth & development, Muscle, Skeletal innervation, Rats, Rats, Inbred Lew, Recovery of Function, Sciatic Nerve injuries, Sciatic Nerve surgery, Stress, Physiological, Nerve Regeneration physiology, Neurons transplantation
- Abstract
Introduction: Acellular nerve allografts (ANAs) yield less consistent favorable outcomes compared with autografts for long gap reconstructions. We evaluated whether a hybrid ANA can improve 6-cm gap reconstruction., Methods: Rat sciatic nerve was transected and repaired with either 6-cm hybrid or control ANAs. Hybrid ANAs were generated using a 1-cm cellular isograft between 2.5-cm ANAs, whereas control ANAs had no isograft. Outcomes were assessed by graft gene and marker expression (n = 4; at 4 weeks) and motor recovery and nerve histology (n = 10; at 20 weeks)., Results: Hybrid ANAs modified graft gene and marker expression and promoted modest axon regeneration across the 6-cm defect compared with control ANA (P < 0.05), but yielded no muscle recovery. Control ANAs had no appreciable axon regeneration across the 6-cm defect., Discussion: A hybrid ANA confers minimal motor recovery benefits for regeneration across long gaps. Clinically, the authors will continue to reconstruct long nerve gaps with autografts. Muscle Nerve 57: 260-267, 2018., (© 2017 Wiley Periodicals, Inc.)
- Published
- 2018
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44. Stem cell transplantation for Huntington's diseases.
- Author
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Choi KA, Choi Y, and Hong S
- Subjects
- Animals, Cell Differentiation genetics, Disease Models, Animal, Humans, Neurons pathology, Neurons transplantation, Huntington Disease therapy, Induced Pluripotent Stem Cells transplantation, Neural Stem Cells transplantation, Stem Cell Transplantation trends
- Abstract
Therapeutic approaches based on stem cells have received considerable attention as potential treatments for Huntington's disease (HD), which is a fatal, inherited neurodegenerative disorder, caused by progressive loss of GABAergic medium spiny neurons (MSNs) in the striatum of the forebrain. Transplantation of stem cells or their derivatives in animal models of HD, efficiently improved functions by replacing the damaged or lost neurons. In particular, neural stem cells (NSCs) for HD treatments have been developed from various sources, such as the brain itself, the pluripotent stem cells (PSCs), and the somatic cells of the HD patients. However, the brain-derived NSCs are difficult to obtain, and the PSCs have to be differentiated into a population of the desired neuronal cells that may cause a risk of tumor formation after transplantation. In contrast, induced NSCs, derived from somatic cells as a new stem cell source for transplantation, are less likely to form tumors. Given that the stem cell transplantation strategy for treatment of HD, as a genetic disease, is to replace the dysfunctional or lost neurons, the correction of mutant genes containing the expanded CAG repeats is essential. In this review, we will describe the methods for obtaining the optimal NSCs for transplantation-based HD treatment and the differentiation conditions for the functional GABAergic MSNs as therapeutic cells. Also, we will discuss the valuable gene correction of the disease stem cells by the CRISPR/Cas9 system for HD treatment., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
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45. Neurotransplantation Therapy and Cerebellar Reserve.
- Author
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Cendelin J, Mitoma H, and Manto M
- Subjects
- Humans, Cerebellar Ataxia surgery, Cerebellum transplantation, Neurons transplantation
- Abstract
Background & Objective: Neurotransplantation has been recently the focus of interest as a promising therapy to substitute lost cerebellar neurons and improve cerebellar ataxias. However, since cell differentiation and synaptic formation are required to obtain a functional circuitry, highly integrated reproduction of cerebellar anatomy is not a simple process. Rather than a genuine replacement, recent studies have shown that grafted cells rescue surviving cells from neurodegeneration by exerting trophic effects, supporting mitochondrial function, modulating neuroinflammation, stimulating endogenous regenerative processes, and facilitating cerebellar compensatory properties thanks to neural plasticity. On the other hand, accumulating clinical evidence suggests that the self-recovery capacity is still preserved even if the cerebellum is affected by a diffuse and progressive pathology. We put forward the period with intact recovery capacity as "restorable stage" and the notion of reversal capacity as "cerebellar reserve"., Conclusion: The concept of cerebellar reserve is particularly relevant, both theoretically and practically, to target recovery of cerebellar deficits by neurotransplantation. Reinforcing the cerebellar reserve and prolonging the restorable stage can be envisioned as future endpoints of neurotransplantation., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)
- Published
- 2018
- Full Text
- View/download PDF
46. Modified acellular nerve-delivering PMSCs improve functional recovery in rats after complete spinal cord transection.
- Author
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Tian T, Yu Z, Zhang N, Chang Y, Zhang Y, Zhang L, Zhou S, Zhang C, Feng G, and Huang F
- Subjects
- Animals, Cell Differentiation genetics, Female, Neurons cytology, Neurons transplantation, Placenta cytology, Placenta transplantation, Pregnancy, Rats, Recovery of Function genetics, Recovery of Function physiology, Spinal Cord growth & development, Spinal Cord transplantation, Spinal Cord Injuries genetics, Spinal Cord Injuries physiopathology, Mesenchymal Stem Cell Transplantation methods, Mesenchymal Stem Cells cytology, Nerve Regeneration genetics, Spinal Cord Injuries therapy, Tissue Engineering
- Abstract
Due to the poor regeneration capacity of neurons and the inhibitory microenvironment, spontaneous regeneration in spinal cord injury (SCI) remains challenging. Tissue engineering is considered a promising approach for enhancing the regeneration of SCI by reconstructing the inherent structure and improving the microenvironment. In this study, the possibility of engineering a nerve complex, which is constructed by acellular nerve delivering placenta mesenchymal stem cells (PMSCs), was assessed for the recovery of a transected spinal cord. Modified acellular nerve grafts were developed, and PMSCs labeled with green fluorescent protein (GFP) were seeded on the graft to construct the engineered nerve complex. Then, the engineered nerve complex was implanted into a 2 mm-length transected gap of the spinal cord. Four weeks after the transplantation, numerous surviving PMSCs were observed in the lesion cavity by immunofluorescence staining. Moreover, co-localization between GFP and neurofilament-200 (NF200) and Neuronal Class III β-Tubulin (Tuj1) was observed at the bridge interface. The PMSCs-graft group exhibited significant function improvement as evaluated by the Basso, Beattie and Bresnahan (BBB) locomotion score and footprint analysis. Eight weeks after surgery, the evoked response was restored in the PMSCs-graft group and numerous thick myelin sheathes were observed compared to that in the control groups. Collectively, our findings suggest that the nerve complex prepared by acellular nerve delivering PMSCs enhanced the structure and function regeneration of the spinal cord after SCI.
- Published
- 2017
- Full Text
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47. The zinc finger E-box-binding homeobox 1 ( Zeb1 ) promotes the conversion of mouse fibroblasts into functional neurons.
- Author
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Yan L, Li Y, Shi Z, Lu X, Ma J, Hu B, Jiao J, and Wang H
- Subjects
- Animals, Basic Helix-Loop-Helix Transcription Factors genetics, Basic Helix-Loop-Helix Transcription Factors metabolism, Cells, Cultured, Embryo, Mammalian cytology, Fibroblasts cytology, Gene Expression Profiling, Germ-Free Life, Hippocampus, Mice, Inbred C57BL, Mice, Inbred ICR, Nerve Tissue Proteins antagonists & inhibitors, Nerve Tissue Proteins genetics, Neurons cytology, Neurons transplantation, POU Domain Factors genetics, POU Domain Factors metabolism, RNA Interference, Recombinant Proteins metabolism, Zinc Finger E-box-Binding Homeobox 1 antagonists & inhibitors, Zinc Finger E-box-Binding Homeobox 1 genetics, Cell Transdifferentiation, Fibroblasts metabolism, Gene Expression Regulation, Developmental, Nerve Tissue Proteins metabolism, Neurogenesis, Neurons metabolism, Zinc Finger E-box-Binding Homeobox 1 metabolism
- Abstract
The zinc finger E-box-binding transcription factor Zeb1 plays a pivotal role in the epithelial-mesenchymal transition. Numerous studies have focused on the molecular mechanisms by which Zeb1 contributes to this process. However, the functions of Zeb1 beyond the epithelial-mesenchymal transition remain largely elusive. Using a transdifferentiation system to convert mouse embryonic fibroblasts (MEFs) into functional neurons via the neuronal transcription factors achaete-scute family bHLH (basic helix-loop-helix) transcription factor1 ( Ascl1 ), POU class 3 homeobox 2 (POU3F2/ Brn2 ), and neurogenin 2 (Neurog2, Ngn2 ) (ABN), we found that Zeb1 was up-regulated during the early stages of transdifferentiation. Knocking down Zeb1 dramatically attenuated the transdifferentiation efficiency, whereas Zeb1 overexpression obviously increased the efficiency of transdifferentiation from MEFs to neurons. Interestingly, Zeb1 improved the transdifferentiation efficiency induced by even a single transcription factor ( e.g. Asc1 or Ngn2 ). Zeb1 also rapidly promoted the maturation of induced neuron cells to functional neurons and improved the formation of neuronal patterns and electrophysiological characteristics. Induced neuron cells could form functional synapse in vivo after transplantation. Genome-wide RNA arrays showed that Zeb1 overexpression up-regulated the expression of neuron-specific genes and down-regulated the expression of epithelial-specific genes during conversion. Taken together, our results reveal a new role for Zeb1 in the transdifferentiation of MEFs into neurons., (© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2017
- Full Text
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48. Direct Reprogramming of Fibroblasts via a Chemically Induced XEN-like State.
- Author
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Li X, Liu D, Ma Y, Du X, Jing J, Wang L, Xie B, Sun D, Sun S, Jin X, Zhang X, Zhao T, Guan J, Yi Z, Lai W, Zheng P, Huang Z, Chang Y, Chai Z, Xu J, and Deng H
- Subjects
- Aging, Animals, Animals, Newborn, Brain cytology, Cell Differentiation, Cell Lineage, Cell Survival, Cells, Cultured, Female, Gene Expression Profiling, Genomic Instability, Green Fluorescent Proteins metabolism, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Male, Mice, Neurons cytology, Neurons metabolism, Neurons transplantation, Transcription, Genetic, Cellular Reprogramming, Endoderm cytology, Extraembryonic Membranes cytology, Fibroblasts metabolism
- Abstract
Direct lineage reprogramming, including with small molecules, has emerged as a promising approach for generating desired cell types. We recently found that during chemical induction of induced pluripotent stem cells (iPSCs) from mouse fibroblasts, cells pass through an extra-embryonic endoderm (XEN)-like state. Here, we show that these chemically induced XEN-like cells can also be induced to directly reprogram into functional neurons, bypassing the pluripotent state. The induced neurons possess neuron-specific expression profiles, form functional synapses in culture, and further mature after transplantation into the adult mouse brain. Using similar principles, we were also able to induce hepatocyte-like cells from the XEN-like cells. Cells in the induced XEN-like state were readily expandable over at least 20 passages and retained genome stability and lineage specification potential. Our study therefore establishes a multifunctional route for chemical lineage reprogramming and may provide a platform for generating a diverse range of cell types via application of this expandable XEN-like state., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
49. First state-approved embryonic stem cell trials in China.
- Subjects
- Cell Differentiation genetics, China, Clinical Trials as Topic, Humans, Parkinson Disease genetics, Parkinson Disease pathology, Embryonic Stem Cells transplantation, Neurons transplantation, Parkinson Disease therapy
- Published
- 2017
- Full Text
- View/download PDF
50. Lineage Reprogramming of Astroglial Cells from Different Origins into Distinct Neuronal Subtypes.
- Author
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Chouchane M, Melo de Farias AR, Moura DMS, Hilscher MM, Schroeder T, Leão RN, and Costa MR
- Subjects
- Animals, Astrocytes metabolism, Basic Helix-Loop-Helix Transcription Factors genetics, Cell Lineage, Cells, Cultured, Cerebral Cortex cytology, Cerebral Cortex metabolism, Cerebral Cortex surgery, Mice, Nerve Tissue Proteins genetics, Neurons metabolism, Neurons transplantation, Transfection, Astrocytes cytology, Cellular Reprogramming, Neurons cytology
- Abstract
Astroglial cells isolated from the rodent postnatal cerebral cortex are particularly susceptible to lineage reprogramming into neurons. However, it remains unknown whether other astroglial populations retain the same potential. Likewise, little is known about the fate of induced neurons (iNs) in vivo. In this study we addressed these questions using two different astroglial populations isolated from the postnatal brain reprogrammed either with Neurogenin-2 (Neurog2) or Achaete scute homolog-1 (Ascl1). We show that cerebellum (CerebAstro) and cerebral cortex astroglia (CtxAstro) generates iNs with distinctive neurochemical and morphological properties. Both astroglial populations contribute iNs to the olfactory bulb following transplantation in the postnatal and adult mouse subventricular zone. However, only CtxAstro transfected with Neurog2 differentiate into pyramidal-like iNs after transplantation in the postnatal cerebral cortex. Altogether, our data indicate that the origin of the astroglial population and transcription factors used for reprogramming, as well as the region of integration, affect the fate of iNs., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
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