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1
Dunphy, Jaclyn Marie. "Infection of Neural Stem Cells with Murine Leukemia Viruses Inhibits Oligodendroglial Differentiation: Implications for Spongiform Neurodegeneration". Kent State University / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=kent1334343584.
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Avola, Rosanna. "Dynamic expression of aquaporins in physiological and pathophysiological in vitro models". Doctoral thesis, Università di Catania, 2017. http://hdl.handle.net/10761/3620.
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Water is the main component of biological fluids and a prerequisite of all organisms living. In 1987, Agre isolated a new integral membrane protein acting as a channel that mediates the water flux and uncharged solutes across biological membranes. This protein was called aquaporin1 and ever since its discovery, more than 300 homologues have been identified in animal, bacteria and plant. In human have been discovered 13 aquaporins (AQPs) isoform (AQP0-AQP12) widely distributed in various epithelia and endothelia where are important actors of fluid homeostasis in secretory and absorptive processes in response to an osmotic or pressure gradient. In the human brain nine aquaporin subtypes (AQP1, 2, 3, 4, 5, 7, 8, 9, and 11) have been recognized and partially characterized, but only three aquaporins (AQP1, 4, and 9) have been clearly identified in vivo. This discovery highlighted the concept of the important role of AQPs in all brain functions and of the dynamics of water molecules in the cerebral cortex. Additionally, AQPs relieved an important role in glial control and neuronal excitability, such as in brain structure and general development. However, a clearer understanding of specific function and distribution of water channels in adult or in development brain requires a more detailed elucidation. Some of these findings are limited from the complexity of direct investigation, inaccessibility of the neural tissue, and hence difficulty in obtaining a brain biopsy, until after the death of an individual. In this sense, several past and present in vitro models have been used to provide important clues about many processes, such as brain development, neurotoxicity, inflammation, pathogenic mechanisms of the diseases and potential pharmacological targets. In the Chapter I, we have reviewed some in vitro approaches used to investigate the mechanisms involved in Krabbe disease with particular regard to the cellular systems employed to study processes of inflammation, apoptosis and angiogenesis. In this study, we used some in vitro methods with the aim to update the knowledge on stem cells biology and to provide a relationship between aquaporins expression and cellular differentiation. In particular, we have analysed the differentiation of human mesenchymal stem cells from adipose tissue (AT-MSCs) into neural phenotypes and SH-SY5Y neuroblastoma cell line into physiological and pathophysiological dopaminergic neurons. In the Chapter II, we have reported the results of the expression of AQP1, 4, 7, 8 and 9 at 0, 14, and 28 days in AT-MSCs during the neural differentiation by immunocytochemistry, RT-PCR and Western blot analysis. Our studies demonstrated that AT-MSCs could be differentiated into neurons, astrocytes and oligodendrocytes, showing reactivity not only for the typical neural markers, but also for specific AQPs in dependence from differentiated cell type. Our data revealed that at 28 days AT-MSCs express AQP1, astrocytes AQP1, 4 and 7, oligodendrocytes AQP1, 4 and 8, and finally neurons AQP1 and 7. In the Chapter III, we have examined the possible involvement of AQPs in a Parkinson s disease-like cell model. For this purpose, we used SH-SY5Y cell line, differentiated in dopaminergic neurons with retinoic acid (RA) and phorbol 12-myristate 13-acetate (MPA) alone or in association. The vulnerability to dopaminergic neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) and H2O2 was evaluated and compared in all cell groups. We found that the vulnerability of cells was linked to dynamic changes of AQP4 and AQP9. The data described here provides fundamental insights on the biology of the human mesenchymal stem cells and significant evidences on the involvement of AQPs in a variety of physiological and pathophysiological processes. This suggests their possible application as markers, which may be helpful in diagnosing as well as in the understanding of neurodegenerative diseases for future therapeutic approaches.
3
Joannides, Alexis. "Neural differentiation of human embryonic stem cells". Thesis, University of Cambridge, 2009. https://www.repository.cam.ac.uk/handle/1810/252121.
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Human embryonic stem cells (hESCs) are a potential source of defined cell types for studying early human development and application in regenerative medicine. Realising this potential requires a number of challenges to be overcome. The experimental findings reported represent a systematic approach in establishing controlled and standardised conditions for differentiating hESCs down the neural lineage, and characterising neural derivatives both in vitro and in vivo. Human embryonic stem cell cultures were established from two independently-derived liens, H9 and UES9. A novel, efficient method for propagating hESCs is described, avoiding the use of enzymatic products which may lead to karyotypic instability. Controlled neuroectodermal differentiation is demonstrated using a chemically defined system over a period of 16 days, and this process is shown to be dependent on endogenous fibroblast growth factor (FGF) signalling. Neural progenitors generated with this system are subsequently expanded for over 180 days and shown to retain neural stem cell (NSC) identity at the clonal level. Evidence is provided that hESC-derived NSCs follow a developmentally predictable timecourse of neurogenesis followed by gliogenesis, and their in vitro and in vivo behaviour is characterised with respect to temporal maturation and phenotypic potential.
4
Eriksson, Malin. "Manipulating neural stem cells". Stockholm, 2010. http://diss.kib.ki.se/2010/978-91-7409-853-2/.
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Pan, Chendong. "Neural differentiation from human embryonal carcinoma stem cells". Thesis, Durham University, 2007. http://etheses.dur.ac.uk/2460/.
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It is understood that retinoic acid (RA), sonic hedgehog (Shh) and bone morphogenic proteins (BMPs) play an important role in cell fate determination and the specification of inter-neurons and motor neurons along the dorsal-ventral axis in the neural tube. In this study, we investigated the function of these signalling molecules to instruct the differentiation of human pluripotent stem cells to form specific neuronal subtypes. TERA2.cl.SP12 embryonal carcinoma (EC) cells are a robust caricature of human embryogenesis and an accepted model of neural differentiation. Gene and protein expression analyses using RT-PCR, western blotting and immunocytochemical techniques indicated that human EC cells respond to RA, BMPs and Shh in a conserved manner and regulate neural transcription factors and structural proteins in a predicted way as cells commit toward the motor neuron phenotype. To assess the function of these differentiated neurons, we tested their ability to innervate skeletal muscle myotubes and induce muscle cell contraction. Myotubes contracted only when cocultured with neurons. The number of contractile events increased significantly when cells differentiated into motor neurons were cocultured with myotubes compared to cocultures with cells that formed intemeurons. Staining for α-bungarotoxin showed positive staining in a pattern characteristic of boutons found in neuromuscular junctions. We also showed that muscle contraction could be manipulated pharmacologically: curare and atropine blocked myotube contraction, whereas acetylcholine and carbachol increased the number of contractile events. In other experiments, we have also shown that cells exposed to RA and Shh in conjunction with other growth factors over different time periods, preferentially form oligodendrocytes and/or interneurons. These results indicate it is feasible to control and direct the differentiation of human stem cells and produce specific neuron subtypes in vitro. Furthermore, this system acts as a useful model to investigate the molecular mechanisms and signalling pathways that control neural differentiation in man.
6
Kennea, Nigel Leonard. "Neural differentiation of human fetal mesenchymal stem cells". Thesis, Imperial College London, 2007. http://hdl.handle.net/10044/1/7409.
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The potential of mesenchymal stem cells (MSC) to differentiate into neural lineages has raised the possibility of autologous cell transplantation as therapy for neurological diseases. There are, however, no studies reporting significant numbers of oligodendrocytes, the myelinforming cells of the central nervous system, derived from MSC. We have recently identified a population of circulating human fetal MSC that are highly proliferative and readily differentiate into bone, cartilage, fat and muscle. I demonstrated for the first time that primary fetal MSC differentiate into cells resemblifl neural precursors and then oligodendrocytes both in vitro and in vivo. By exposing cells to a neuronal conditioned medium, rates of oligodendrocyte differentiation approaching 50% were observed, and cells appeared to mature appropriately in culture. Importantly, the differentiation of a clonal population into both mesodermal (bone) and ectodermal (oligodendrocyte) lineages was achieved. In the developing murine brain, cells integrated but oligodendrocyte differentiation of naiVe fetal MSC was very low. The proportion of oligodendrocyte differentiation was increased (from 0.2% to 4%) by pre-exposing the cells to differentiation medium prior to transplantation. The process of in vivo differentiation occurred without cell fusion. Although the main focus of this thesis was oligodendrocyte differentiation, I also recapitulated controversial published work into neuronal differentiation of MSC. The exposure of cells to the reducing agent butylated hydroxyanisole induced rapid changes in cell morphology and expression of neuronal markers. These 'differentiated' cells did not, however, appear functional with no upregulation of voltage-gated sodium channels or synaptophysin. Finally, while stem cells offer promise for correction of brain diseases, one major obstacle is the poor survival of grafted cells. Investigation of apoptotic signalling showed fetal MSC have functional apoptotic machinery in both the intrinsic (mitochondrial) and extrinsic (death receptor) pathways which could be manipulated to prolong stem cell survival by inhibition of death signalling.
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Erlandsson, Anna. "Neural Stem Cell Differentiation and Migration". Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl.[distributör], 2003. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3546.
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Hardy, Steven Allan. "Mesenchymal stem cells as trophic mediators of neural differentiation". Thesis, Durham University, 2010. http://etheses.dur.ac.uk/524/.
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Intense excitement and optimism surrounds the rapidly-expanding field of stem cell research, owing to their high capacity for self-renewal and intrinsic ability to differentiate into mature cell lineages. Although it may be envisioned that embryonic stem cells will be of significantly greater therapeutic value than their adult stem cell counterparts, the use of embryonic stem cells is fraught with both technical and ethical challenges and, as such, significant impetus has been placed on adult stem cell-based research. In particular, mesenchymal stem cells (MSCs) present as exciting candidates for potential use in cellular therapies and tissue engineering strategies. MSCs are defined at the functional level in terms of their ability to differentiate into mesodermal derivatives such as bone and fat. However, this functional definition is evolving, and there is considerable evidence to suggest that MSCs have a key role within their niche involving the release and/or uptake of soluble factors and cytokines, significantly influencing the behaviour of other cell types within the niche. Both facets of MSC behaviour are valuable from a clinical perspective, and have been examined in the present thesis. The most obvious and realistically-achievable clinical application of MSCs at present is in the treatment of osseous and adipose tissue defects. However, before the use of MSCs in the clinic becomes more commonplace, it is crucial to gain a more comprehensive understanding of the complex molecular and cellular mechanism(s) by which MSCs commit to a given fate and undergo differentiation to produce mature, fully-functional derivatives. Much of our present knowledge is derived from studies performed on the highly unnatural, 2D environment of tissue culture plastic. The present study assessed the behaviour of MSCs cultured on AlvetexTM, a novel, 3D scaffold manufactured by ReInnervate, with particular emphasis on the ability of MSCs to undergo osteogenic and adipogenic differentiation. Results obtained suggest that AlvetexTM may provide a more realistic and physiologically-relevant system in which to study osteogenesis and adipogenesis, in a manner more pertinent to that which occurs in vivo. Furthermore, the ability of MSCs to influence the behaviour of other cell types via the release of trophic factors and cytokines was examined, with particular emphasis on the nervous system. An in vitro conditioned media model was developed in order to investigate the influence(s) of MSC-derived soluble factors/cytokines on neural development and plasticity, using the adult rat hippocampal progenitor cell (AHPC) line as a model system. Results obtained suggest that, under defined conditions, MSCs secreted a complement of soluble factors/cytokines that induce AHPCs to commit to and undergo astrogenesis. This effect was characterised at both the cellular and molecular level. The specific complement of bioactive factors secreted by MSCs has been investigated using a combination of targeted transcriptional profiling and shotgun proteomics, and several putative candidate factors have been identified for further investigation.
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Albertson, Roger Joseph. "Establishing asymmetry in Drosophila neural stem cells /". view abstract or download file of text, 2003. http://wwwlib.umi.com/cr/uoregon/fullcit?p3112998.
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Thesis (Ph. D.)--University of Oregon, 2003.Typescript. Includes vita and abstract. Includes bibliographical references (leaves 101-117). Also available for download via the World Wide Web; free to University of Oregon users.
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Jones, Robert. "Proteomic analysis of neural differentiation in mouse embryonic stem cells". Thesis, Bangor University, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.412699.
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Wiskow, Ole. "Evaluation of the neuronal differentiation capacity of pluripotent stem cells and neural stem cells in monolayer culture". Thesis, University of Cambridge, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.609417.
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EL, SAID DALYA. "Blood derived stem cells (BDSCs): neural differentiation protocols for human therapy". Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2012. http://hdl.handle.net/2108/209984.
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La tecnologia delle cellule staminali hanno causato un notevole entusiasmo tra gli interessati alla salute degli animali e degli uomini. Molte persone stanno cercando nuove cure per le malattie e la terapia i vivo. Noi sappiamo che tutti i tipi di cellule possono essere rigenerate eccetto le cellule neuronali di mammifero, sebbene il tessuto nervoso periferico si rigeneri la guaina mielinica che permette l’orientamento della fibra. L’importanza di ottenere cellule neuronali funzionali è vitale per le malattie neurodegenerative. Le malattie neurodegenerative sono per definizione delle malattie progressive corniche caratterizzate da una selettiva perdita di neuroni in aree motori simmetriche, sensoriali e cognitive appartenenti al SNC, o perdita/ disfunzione delle fibre mieliniche o non mieliniche nel SNP. Oggi è possibile deprogrammare cellule adulte in cellule staminali pluripotenti in vitro utilizzando specifici protocolli e sostanze non tossiche per l’uomo, ottenere neurosfere (corpi cellulari da dove è possibile isolare e studiare le cellule staminali neuronali). In questo lavoro di tesi ho dimostrato come è possibile utilizzando cellule deprogrammate del sangue ottenere neurosfere e differenziarle in neuroni adulti in vitro e come questi miei risultati siano in accordo con i risultati ottenuti in vivo.Stem cells technology has provoked considerable excitement among people interested in welfare of animals and humans. Many people are searching for new treatments of diseases and in vivo therapy. We know that all types of cells can be regenerated except neural cells of mammals such as don’t regenerate, although the peripheral nervous tissue regenerate if neurillematic sheath allow the orientation of fiber. the importance of obtaining functional nerve cells is vital in neurodegenerative diseases. Neurodegenerative diseases are by definition progressive chronic diseases characterized by a selective loss of neurons in areas, symmetric motor , sensory , cognitive and member ship of CNS, or loss / dysfunction of myelinated or non myelinated fibers in the PNS. It has been possible today to use BDSCs after Deprogrammation of adult peripheral blood, and using specific protocols to obtain “neurospheres” from these cells in specific medium and non toxic substances addition. Neurospheres (composed of neural stem cells) provide a method for investigating neural precursor in vitro instead of embryonic stem cells. In this study I have demonstrated how BDSCs being able to form neurospheres and differentiated into mature neurons in vitro, and how it is possible to use stem cells therapy as treatment in vivo.
13
Sartor, Francesca. "Regulation of translation initiation and RNA decay is important for neuronal differentiation". Thesis, University of Aberdeen, 2016. http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=.
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Ge, Shufan. "Impact of Muscarinic Receptor Activation on Neural Stem Cell Differentiation". University of Toledo Health Science Campus / OhioLINK, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=mco1291827018.
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Nordin, Norshariza. "Roles of Wnt signalling during neural differentiation of embryonic stem (ES) cells". Thesis, University of Edinburgh, 2006. http://hdl.handle.net/1842/25031.
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In order to investigate how Wnt signalling affects the differentiation of ES cells into neurons in vitro. RNA expression of all 19 Wnt genes together with two Wnt antagonists, Dkk1 and sFRP2, during the process was first determined by RT-PCR. Of 19 Wnt genes, the expression of 12 with particularly interesting patterns was subsequently analysed by quantitative RT-PCR (qRT-PCR). Neural differentiation of ES cells was induced through the formation of embryoid bodies (EBs) and the addition of retinoic acid (RA). The gene expression was determined in five different stages of the process including undifferentiated ES cells, early EBs prior to the addition of RA, EBs in the presence of RA, attached EBs after withdrawal of RA and finally neuron-like cells grown on an adhesive substratum. Many Wnt genes showed dynamic alterations in expression levels during neural differentiation. In order to test the effect of expressing Wnts at different stages, we developed an inducible expression system. The system is based on cre-loxP strategy and allows for stimulation or inhibition of Wnt signalling at specific steps of the differentiation process. Overexpression of Wnt1-HA and Wnt3a at early and late stages of the process in addition to constitutive expression of these genes was carried out. Additionally, inhibition of Wnt signalling by overexpressing Dkk1 at early and late stages as well as its constitutive expression during the differentiation process was also conducted. The effects of altered Wnt signalling on the formation of neural precursor cells (npc) and neuronal cells was analysed using specific protein markers by Immunocytochemistry and fluorescent-activated cell sorting (FACS) analysis. It was found that overexpression of Wnt1 and Wnt3a at the early stages of the differentiation process significantly reduced the formation of npc while inhibition of Wnt signalling at this stage by induction of Dkk1 significantly increased the number of npc. Interestingly, constitutive expression of each of these genes reduced the number of npc. Formation of neurons was stimulated by overexpression of Wnt3a at late stages of neural differentiation process, whereas induction of Wnt1 at this stage enhanced the percentage of neurons in cultures. Overexpression of Dkk1 at this stage however significantly decreased the formation of npc. A significant increase in the percentage of neuron formation was observed when Dkk1 and Wnt1 were overexpressed at early stages of the process. In contrast, overexpression of Wnt3a at this stage significantly reduced the number of neurons. Additionally, constitutive stimulation or inhibition of Wnt signalling during the process was observed to reduce the ability of ES to differentiate into neurons. These observations therefore highlight the complexity of probable function of stage-dependent response to Wnt signalling during neural differentiation.
16
Yuen, Shun Ming. "Regulation of neural differentiation in mouse embryonic stem cells using small molecules". Thesis, Cardiff University, 2013. http://orca.cf.ac.uk/53933/.
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Embryonic stem (ES) cells are a potential source of neural derivatives that can be used in stem cell-based therapies. To generate specific cell types in a predictable manner, a detailed understanding of cell fate specification is required. To address this, this study employed mouse ES cells as a model to explore how neural identity was acquired and how regional identities were specified in ES cell-derived neural progenitor cells (NPCs). Chemical inhibitors are more stable than recombinant proteins, hence they give more reproducible biological responses. This is crucial when generating specific cell types from ES cells for large-scale analysis. This study explored the effect of two newly discovered small molecule inhibitors of BMP signalling, dorsomorphin (DM) and LDN193189 (LDN) on the neural induction of ES cells, and compared their capabilities with those of recombinant noggin. Both DM and LDN treatments increased the expression of neural markers to levels that were comparable to that achieved by noggin treatment, suggesting that both LDN and DM can potentially substitute recombinant noggin in the generation of NPCs in vitro. Retinoic acid (RA) is an important regulator of regional specification in vivo, but the underlying mechanisms for giving region-specific response are unclear. Here, early and late NPCs were exposed to RA at specific periods and were analysed for neural and positional markers. Results showed that region-specific responses were produced by RA at specific periods, indicating that NPCs display temporal changes in patterning responsiveness to morphogenic cues. The ability of morphogens to impose positional response is lost in late cultures. Since Sox1 expression is associated with a naïve neural progenitor state and neural commitment, the expression of Sox1 was examined over time to test whether it was associated with the responsiveness to patterning cues. Both caudal and ventral markers were induced by RA and, because it plays an essential role in ventral patterning, Hedgehog (Hh) agonist purmorphamine respectively in early NPCs but not in late NPCs in both Sox1 positive and negative populations. This suggested that the expression of positional markers was not dependent on a temporally-defined Sox1 progenitor state.
17
Liu, Ning. "Expansion and Neural Differentiation of Embryonic Stem Cells in Three-Dimensional Cultures". The Ohio State University, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=osu1262281522.
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Joshi, Ramila Joshi. "Micro-engineering of embryonic stem cells niche to regulate neural cell differentiation". University of Akron / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=akron1544029342969082.
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Doszyn, Olga. "Sex differences in neuronal differentiation of human stem cells". Thesis, Uppsala universitet, Institutionen för biologisk grundutbildning, 2019. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-384661.
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Sexual dimorphism has been long noted in human neurobiology, apparent most notably in sex-biased distribution of multiple neurological disorders or diseases, from autism spectrum disorder to Parkinson's disease. With the advances in molecular biology, genetics and epigenetics have come into focus as key players in sexually dimorphic neural development; and yet, many studies in the field of neuroscience overlook the importance of sex for the human brain. For this project, human embryonic and neural stem cells were chosen for three main reasons. Firstly, they provide an easily obtainable, scalable and physiologically native model for the early stages of development. Secondly, neural stem cells populations are retained within the adult human brain, and are implicated to play a role in cognition and mental illness, and as such are of interest in themselves. Thirdly, stem cell lines are widely used in research, including clinical trials of transplantation treatments, and for this reason should be meticulously examined and characterized. Here, the morphology, behaviour, and expression of selected genes in four stem cell lines, two of female and two of male origin, was examined in side-by-side comparisons prior to and during neuronal differentiation using a variety of methods including light microscopy, time-lapse two-photon microscopy, quantitative real-time PCR and immunocytochemistry. The obtained results have shown previously uncharacterised differences between those cell lines, such as a higher rate of proliferation but a slower rate of neuronal differentiation in male cell cultures compared to female cells cultivated in the same conditions, and a sex-biased expression of several markers of neuronal maturation at late stages of differentiation, as well as diverse patterns of expression of X- and Y-linked genes involved in stem cell proliferation and neural development.
20
Enarsson, Mia. "Roles of PDGF for Neural Stem Cells". Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2004. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-4245.
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Aarum, Johan. "Interactions between mouse CNS cells: microglia and neural precursor cells /". Stockholm, 2004. http://diss.kib.ki.se/2004/91-7140-120-2/.
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Kato, Takeo. "A neurosphere-derived factor, cystatin C, supports differentiation of ES cells into neural stem cells". Kyoto University, 2006. http://hdl.handle.net/2433/135642.
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Chabu, Chiswili Yves. "Regulation of cell polarity and self-renewal in Drosophila neural stem cells /". Connect to title online (Scholars' Bank) Connect to title online (ProQuest), 2008. http://hdl.handle.net/1794/8330.
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Thesis (Ph. D.)--University of Oregon, 2008.Typescript. Includes vita and abstract. Includes bibliographical references (leaves 82-93). Also available online in Scholars' Bank; and in ProQuest, free to University of Oregon users.
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Faijerson, Jonas. "Neural stem/progenitor cells in the post-ischemic environment : proliferation, differentiation and neuroprotection /". Göteborg : Center for Brain Repair and Rehabilitation, Institute of Neuroscience and Physiology, Göteborg University, 2007. http://hdl.handle.net/2077/4516.
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Nasir, Wafaa. "Effect of Topography on Mouse Embryonic Stem Cells During Pluripotency and Neural Differentiation". University of Akron / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=akron1533098386697352.
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Holmström, Niklas. "Directed differentiation of adult neural stem cells for cell therapy in the nervous system /". Stockholm, 2005. http://diss.kib.ki.se/2005/91-7140-358-2/.
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Ovando, Roche Patrick. "Role of telomere binding protein TRF2 in neural differentiation of human embryonic stem cells". Thesis, Imperial College London, 2013. http://hdl.handle.net/10044/1/24848.
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Telomere repeat binding factor 2 (TRF2) is reported to be a key component of shelterin, a multi-protein complex that binds telomeric deoxyribonucleic acid (DNA) to protect chromosome ends and maintain genome stability. However, in recent years, TRF2 has been found to also bind non-telomeric regions and to act as a protein hub, interacting with a wide range of non-telomeric proteins and thus raising the possibility that it may serve functions independent of telomere maintenance. Despite the importance of TRF2, there is little information about how TRF2 is expressed during development and whether it could have an extratelomeric role in this process. Human embryonic stem cells (hESCs) derived from pre-implantation embryos are able to differentiate into most, if not all, tissues of the adult body, thereby provide a good cell model to tackle the problem. Given the abundance of TRF2 in the human brain and its potential for extratelomeric roles, this study focused on neural differentiation. TRF2 protein levels were found dramatically increased upon differentiation of hESCs to neural progenitor cells (NPCs) and these high levels, similarly to what is observed in vivo, were specific to the neural lineage. Gain and loss of function approaches revealed that exogenous expression of TRF2 in hESCs induced neural differentiation while, in contrast, TRF2 knockdown in NPCs drastically hindered their ability to terminally differentiate into neurons and glia. This enhancing neural function of TRF2 is achieved through the ability of TRF2 to inhibit the proteasomal degradation of REST4, an alternative splice variant of RE1-Silencing Transcription factor (REST), which alleviates REST repression over neural genes, hence consolidating neural progenitor identity and potency. This study identifies TRF2 as a novel component of neural differentiation, suggesting its importance in central nervous system development as well as in neurological disorders.
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Kishi, Yo. "Estrogen promotes differentiation and survival of dopaminergic neurons derived from human neural stem cells". Kyoto University, 2005. http://hdl.handle.net/2433/144382.
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Kyoto University (京都大学)0048
新制・課程博士
博士(医学)
甲第11935号
医博第2917号
新制||医||910(附属図書館)
23724
UT51-2006-B114
京都大学大学院医学研究科脳統御医科学系専攻
(主査)教授 山中 伸弥, 教授 福山 秀直, 教授 篠原 隆司
学位規則第4条第1項該当
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Repele, Andrea. "Differentiation potential and metabolic analysis of satellite cells and amniotic fluid stem cells". Doctoral thesis, Università degli studi di Padova, 2012. http://hdl.handle.net/11577/3422458.
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We have recently characterized two distinct populations of Satellite Cells (SCs), defined as Low Proliferative Clones (LPC) and High Proliferative Clones (HPC), that differ for proliferation, egenerative potential and mitochondrial coupling efficiency. In here, we have deep investigated their cell biology and characterized features that remark their intrinsic differences retrievable also at the initial phases of their cloning. LPC and HPC can indeed be istinguished for characteristic mitochondrial membrane potential (ΔΨm) just after isolation from their parental fibre. This is merged by mitochondrial redox state measured via NAD+/NADH analysis- and alternative respiratory CO2 production in cloned cells, which are accountable for metabolic differences reflected by alternative expression of the glycolytic enzyme Pfkfb3. In addition also mitochondrial Ca2+ handling and the sensitivity to apoptosis triggered via the intrinsic pathway are modified as well as the size of the mitochondrial network. In conclusion, we were able to determine which clone represents the suitable stem cell within the SCs population. These further experimental observations report novel
physiological features in the cell biology of SCs populations before and after cloning, highlighting an intrinsic heterogeneity on which the stemness of the satellite cell is likely to depend.
In the second part of my work we have also investigated their potential to trans-differentiate into smooth muscle cells. Enteric Nervous System normally interacts with muscle cells to control the peristaltic and secretory activity of the gut wall. Incomplete gut colonization by neural crest cells causes Hirschsprung’s disease, characterized by aganglionosis of the distal bowel. Multipotent, self-renewing enteric precursor neurosphere-like bodies (NLBs) -capable of generating neurons and glia derived from the neural crest- can be isolated from the gut of mice, rats, and human and they are able to colonize the gut after transplantation. Our aim is to understand the relationship between satellite cells-derived muscle precursor cells (MPCs) and NLBs using an in vitro co-culture model: this will be useful in perspective of a tissue engineering approach for bowel regeneration and skeletal muscle. Our records highlighted that NLBs were able to form new myotubes in presence of MPCs. Co-cultures in myogenic medium showed a remarkable improvement of MPCs ifferentiation by NLBs, promoting the formation of sarcomeric striatures onto myotubes and increasing the desmin expression of MPCs. On the other side, using neurogenic medium MPCs-NLBs showed a neural-like phenotype. As future perspectives, we need to understand the relationship between MPCs and NLBs and if the synapses are involved in this process; to verify if the seeding on a biocompatible polymer influences the behaviour of neural cells; and we must confirm these data with an in vivo skeletal and smooth muscle differentiation.
We have finally explored the possibility of deriving smooth muscle cells from a different source, taking in consideration the difficulties related to the expansion of both skeletal and smooth muscle progenitors. Therefore, we aim to derive functional smooth muscle cells (SMCs) from non-muscle cells, such as human Amniotic Fluid Stem (hAFSC) cells. hAFSC were transduced using vector encoding ZsGreen under the αSMA promoter. SMhAFSC expressed significantly higher level of smooth muscle genes (such as αSMA, desmin, calponin and smoothelin expression) after selective culture condition. These features were confirmed by immunofluorescence, demonstrating a single lineage commitment; TEM established increased
intermediate filaments, dense bodies and glycogen deposits in SMhAFSC, similar pattern compared to SMCs; and sequential imaging analyses demonstrated that SMhAFSC have a higher contractile potential than hAFSC. Consecutive single cell sampling showed the presence of voltagedependent calcium activated potassium channels on differentiated SMhAFSC and showed a higher production of carbon dioxide. In conclusion, we were able to generate to functional SMCs starting from a non-muscle precursor; secondly the transduction process may represent a valuable tool to select SM committed population. This step may eventually overcome the well-known problem of expanding SM progenitors, making these cells amenable to tissue engineering.Il nostro gruppo ha recentemente caratterizzato due distinte popolazioni di cellule satelliti, classificate come cloni a bassa proliferazione (LPC) e ad alta proliferazione (HPC), che si differenziano in termini di proliferazione, potenziale rigenerativo e metabolismo mitocondriale. Nel mio lavoro di dottorato, abbiamo valutato e caratterizzato la loro biologia cellulare con particolare attenzione a quelle differenze intrinseche presenti anche prima della loro clonazione. Infatti, ambo le tipologie clonali possono essere distinte mediante il potenziale di membrana mitocondriale (ΔΨm) subito dopo l’isolamento dalla fibra. Questo dato è in accordo con lo stato ossido riduttivo mitocondriale misurato tramite NAD+/NADH e la quantificazione della produzione di CO2. Questi risultati sono responsabili delle differenze metaboliche e possono essere spiegati dalla diversa espressione dell’enzima glicolitico Pfkfb3. Inoltre la concentrazione mitocondriale del Ca2+ e la sensibilità all’apoptosi sono modificate così come la dimensione della rete mitocondriale. In conclusione, siamo stati in grado di determinare quale clone rappresenta la cellula staminale all’interno della popolazione di cellule satelliti. Queste nuove osservazioni sperimentali rivelano caratteristiche fisiologiche della biologia delle popolazioni delle cellule satelliti prima e dopo la clonazione, mettendo in luce un’eterogeneità intrinseca della cellula satellite. Nella seconda parte della mia tesi abbiamo esplorato la possibilità che le cellule satelliti possano, se opportunamente stimolate, trans-differenziarsi in cellule muscolari lisce. Il sistema nervoso enterico normalmente interagisce con le cellule muscolari per controllare l’attività peristaltica e secretoria della parete intestinale. L’incompleta colonizzazione dell’intestino da parte delle cellule della cresta neurale provoca la malattia di Hirschsprung, caratterizzata da aganglionosi del colon distale. Le neurosfere (NLBs), precursori enterici in grado di auto-rinnovarsi, possono generare neuroni e glia; essere isolate dall’intestino di topi, ratti e umani e sono in grado di colonizzare l'intestino dopo il trapianto. Il nostro obiettivo è di capire la relazione tra i precursori di cellule satelliti (MPCs) e NLBs utilizzando un modello in vitro di co-coltura: questo sarà utile in prospettiva di un approccio di ingegneria tissutale per la rigenerazione intestinale e muscolo scheletrico. I nostri dati hanno evidenziato che NLBs, in presenza di MPCs, sono in grado di formare nuovi miotubi. L’uso di terreni di coltura miogenici ha evidenziato un notevole aumento della differenziazione in senso muscolare, promuovendo la formazione di striature ed aumentando l’espressione di desmina. Dall’altra parte, l’utilizzo di terreni di coltura neurogenici ha mostrato un fenotipo simil neurale. Come prospettive future, dobbiamo comprendere ulteriormente la relazione tra MPCs e NLBs e se le sinapsi sono coinvolte in questo processo; si deve verificare se un loro utilizzo su polimeri biocompatibili ne possa influenzare il comportamento, ed infine è necessaria una conferma dei suddetti dati tramite un’analisi di differenziazione in vivo in muscolo scheletrico e liscio. Nella terza ed ultima fase del mio lavoro, ci siamo focalizzati ad esplorare la possibilità che cellule non-muscolari possano, se opportunamente stimolate, differenziare in senso muscolare liscio. Il nostro obiettivo è stato quello di ottenere cellule muscolari lisce (SMCs) partendo da cellule staminali del fluido amniotico umano (hAFSC). hAFSC sono state trasdotte utilizzando un virus codificante per ZsGreen sotto il promotore αSMA. SMhAFSC così ottenute hanno evidenziate un alto livello d’espressione dei geni del muscolo liscio (come αSMA, desmina, calponina e smoothelin). Queste caratteristiche sono state confermate da molteplici analisi: di immunofluorescenza, dimostrando la positività a marcatori specifici per il muscolo liscio; microscopia a trasmissione elettronica (TEM), dove si verificava l’aumento della presenza di filamenti intermedi, di corpi densi e depositi di glicogeno, modello simile rispetto alle SMCs. Analisi in timelapse di SMhAFSC hanno dimostrato che queste possiedono un potenziale contrattile superiore rispetto hAFSC e studi su singola cellula hanno evidenziato la presenza di canali calcio voltaggio-dipendenti attivati da potassio solamente su SMhAFSC. In conclusione, siamo stati in grado di generare di cellule muscolari lisce funzionali da un precursore nonmuscolare ed in secondo luogo il processo di trasduzione può rappresentare un valido strumento per distinguere e selezionare differenti popolazioni. Questa fase può eventualmente superare il ben noto problema dell’espansione di progenitori di cellule muscolari lisce, rendendo queste cellule suscettibili per approcci d’ingegneria tessutale.
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Tamm, Christoffer. "Apoptotic cell death in neural stem cells exposed to toxic stimuli /". Stockholm : Karolinska institutet, 2007. http://diss.kib.ki.se/2007/978-91-7357-301-6/.
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Li, Yue y 李越. "Caveolin-1 is a negative regulator of neuronal differentiation of neural progenitor cells in vitro and in vivo". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B46918863.
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Larsson, Jimmy. "Neural stem and progenitor cells cellular responses to known and novel factors /". Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2010. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-110722.
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Contreras-Sesvold, Carmen Sesvold Carmen Contreras. "Reactive astrocytes : phenotypic and functional characteristics and astrocytes as neural stem cells /". Download the thesis in PDF, 2006. http://www.lrc.usuhs.mil/dissertations/pdf/ContrerasSesvold2006.pdf.
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Marzec-Schmidt, Katarzyna. "Deep convolutional neural networks accurately predict the differentiation status of human induced pluripotent stem cells". Thesis, Högskolan i Skövde, Institutionen för biovetenskap, 2020. http://urn.kb.se/resolve?urn=urn:nbn:se:his:diva-19420.
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Rapid progress of AI technology in the life science area is observed in recent years. Convolutionalneural network (CNN) models were successfully applied for the localization and classification of cellson microscopic images. Induced pluripotent stem cells are one of the most important innovations inbiomedical research and are widely used, e.g. in regenerative medicine, drug screening, and diseasemodeling. However, assessment of cell cultures’ quality requires trained personnel, is timeconsumingand hence expensive. Fluorescence microscope images of human induced pluripotentstem‐hepatocytes (hiPS‐HEPs) derived from three human induced pluripotent stem cell (hiPSC) lineswere taken daily from day 1 until day 22 of differentiation. The cells from day 1 to 14 were classifiedas ´Early differentiation´, and above day 16 as ´Late differentiation´. In this study, it wasdemonstrated that a CNN‐based model can be trained with simple fluorescence microscope imagesof human induced pluripotent stem‐hepatocytes, and then used to predict with high accuracy(96.4%) the differentiation stage of an independent new set of images.
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Nishida, Akihiro. "Incorporation and Differentiation of Hippocampus-Derived Neural Stem Cells Transplanted in Injured Adult Rat Retina". Kyoto University, 2001. http://hdl.handle.net/2433/151451.
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Zimmer, Bastian [Verfasser]. "Modeling of neural differentiation by using embryonic stem cells to detect developmental toxicants / Bastian Zimmer". Konstanz : Bibliothek der Universität Konstanz, 2011. http://d-nb.info/1045154121/34.
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Chan, Yan-ho y 陳恩浩. "The influences of lead ions on viability, proliferation and neuronal differentiation of hippocampal-derived neural stem cells of newbornand adult rats". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hub.hku.hk/bib/B48333220.
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Neural stem cells (NSCs) are defined as multipotent stem cells. They are able to self-renew and differentiate into mature cells, such as neurons, oligodendrocytes and astrocytes. Neurotoxicity of lead (Pb2+) has been extensively investigated by many previous studies. These studies proved that lead is a potent toxin that affects nervous system, especially children’s brain. However, most of these studies focused on the negative effects of lead on the differentiated or mature cell types in the brains instead of NSCs. The aim of this study was to reveal the effects of Pb2+ on viability, proliferation and differentiation of NSCs derived from the hippocampus of newborn rats aged 7 days and adult rats aged 90 days in vitro. NSCs harvested from the rat hippocampus were cultured in proliferation medium. After 6-8 days, free-floating neurospheres formed. The neurospheres were dissociated and plated onto poly-L-lysine coated 96-well plate and coverslips. Some dissociated cells were characterized by being stained with anti-nestin to show the presence of NSCs. This project was divided into three parts. In the first part, the Passage 2 (P2) cells plated onto 96-well plate were cultured in the proliferation medium with different concentrations of lead acetate (0-200μM) for 48 hours, followed by 3- (4,5-cimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to detect the effects of Pb2+ on the cell viability. In the second part, P2-NSCs plated onto coverslips in wells were cultured in the proliferation medium with different concentrations of lead acetate (0-200μM). Then, 10 μM bromodeoxyuridine (BrdU) was added into the culture medium for additional 24 hours, followed by immunocytochemistry staining with anti-BrdU. In the last part, the dissociated P2-NSCs plated onto coverslips were allowed to grow in the differentiation medium of neurons, astrocytes or oligodendrocytes with different concentrations of lead acetate (0-200μM). After 6 days, immunocytochemistry staining with anti-microtubule-associated protein 2 (anti-MAP2), anti-glial fibrillary acidic protein (anti-GFAP) or anti-RIP was used to detect the differentiation commitment of affected NSCs.
Low level of Pb2+ (1-10μM) had no effect on the viability of adult hippocampal neural stem cells (hNSCs). However, Pb2+ exposure at the concentration of 10μM could lead to significant cell death of newborn hNSCs. High level of Pb2+ (50-200μM) caused significant cell death of both newborn and adult hNSCs. Newborn hNSCs were sensitive to Pb2+ toxicity in proliferation assay. Even a low concentration (1μM) of lead could lead to significant inhibition of cell proliferation. High level of Pb2+ (50-200μM) suppressed proliferation of both newborn and adult hNSCs significantly. Moderate to high levels of Pb2+ exposure (50-200μM) significant decreased the percentage of mature neurons cultured from both newborn and adult hNSCs. Furthermore, 10μM or more Pb2+could significantly inhibited the oligodendrocyte differentiation of both newborn and adult hNSCs. However, Pb2+ could also stimulate the astrocyte differentiation of hNSCs. Lead concentrations higher than 10μM and 50μM could respectively lead to a significant increase in the percentage of mature astrocytes differentiated from newborn and adult hNSCs. The data showed that Pb2+ inhibited not only the viability and proliferation of rat hNSCs but also the neuronal and oligodendrocyte differentiation in vitro; moreover activated astrocyte differentiation of the hNSCs of both newborn and adult rats were observed with high concentration of Pb2+ in vitro. Also, it was revealed that the hNSCs of newborn rats were more sensitive than those from adult rats to Pb2+ cytoxicity.published_or_final_version
Anatomy
Master
Master of Medical Sciences
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Ngo, Justine Marie. "Understanding Dishevelled-Mediated Wnt Signaling in Regulating Early Development and Stem Cell Differentiation". Case Western Reserve University School of Graduate Studies / OhioLINK, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=case158022929074044.
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Padam, Amith Chordia. "Development and Commercialization of Remyelination Therapeutics to Restore Neural Function in Multiple Sclerosis". Case Western Reserve University School of Graduate Studies / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=case1304690351.
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Phillips, Nick. "Modelling and analysis of oscillations in gene expression through neural development". Thesis, University of Manchester, 2016. https://www.research.manchester.ac.uk/portal/en/theses/modelling-and-analysis-of-oscillations-in-gene-expression-through-neural-development(099f8bee-c1ce-4ca2-951e-a1e3fb7321bd).html.
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The timing of differentiation underlies the development of any organ system. In neural development, the expression of the transcription factor Hes1 has been shown to be oscillatory in neural progenitors, but at a low steady state in differentiated neurons. This change in the dynamics of expression marks the timing of differentiation. We previously constructed a mathematical model to test the experimental hypothesis that the topology of the miR-9/Hes1 network and specifically the accumulation of the micro-RNA, miR-9, could terminate Hes1 oscillations and account for the timing of neuronal differentiation, using deterministic delay differential equations. However, biochemical reactions are the result of random encounters between discrete numbers of molecules, and some of these molecules may be present at low numbers. The finite number of molecules interacting within the system leads to inherent randomness, and this is known as intrinsic stochasticity. The stochastic model predicts that low molecular number causes the time to differentiation to be distributed, which is in agreement with recent experimental evidence and considered important to generate cell type diversity. For the exact same model, fewer reacting molecules causes a decrease in the average time to differentiation, showing that the number of molecules can systematically change the timing of differentiation. Oscillations are important for a wide range of biological processes, but current methods for discovering oscillatory genes have primarily been designed for measurements performed on a population of cells. We introduce a new approach for analysing biological time series data designed for cases where the underlying dynamics of gene expression is inherently noisy at a single cell level. Our analysis method combines mechanistic stochastic modelling with the powerful methods of Bayesian nonparametric regression, and can distinguish oscillatory expression in single cell data from random fluctuations of nonoscillatory gene expression, despite peak-to-peak variability in period and amplitude of single cell oscillations. Models of gene expression commonly involve delayed biological processes, but the combination of stochasticity, delay and nonlinearity lead to emergent dynamics that are not understood at a theoretical level. We develop a theory to explain these effects, and apply it to a simple model of gene regulation. The new theory can account for long time-scale dynamics and nonlinear character of the system that emerge when the number of interacting molecules becomes low. Both the absolute length and the uncertainty in the delay time are shown to be crucial in controlling the magnitude of nonlinear effects.
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Nakaji(Hirabayashi), Tadashi. "Design of Bioactive Materials with Chimeric Proteins for Controlling Proliferation and Differentiation of Neural Stem Cells". 京都大学 (Kyoto University), 2009. http://hdl.handle.net/2433/124507.
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Drury-Stewart, Danielle Nicole. "Controlling the microenvironment of human embryonic stem cells: maintenance, neuronal differentiation, and function after transplantation". Diss., Georgia Institute of Technology, 2011. http://hdl.handle.net/1853/45967.
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Precise control of stem cell fate is a fundamental issue in the use of human embryonic stem (hES) cells in the context of cell therapy We examined three ways in which the microenvironment can be controlled to alter hES cell behavior, providing insight into the best conditions for maintenance of pluripotency and neural differentiation in developmental and therapeutic studies. We first examined the effects of polydimethylsiloxane (PDMS) growth surfaces on hES cell survival and maintenance of pluripotency. Lightly cured, untreated PDMS was shown to be a poor growth surface for hES cells. Some of the adverse effects caused by PDMS could be mitigated with increased curing or UV treatment of the surface, but neither modification provided a growth surface that supported pluripotent hES cells as well as polystyrene. This work provides a basis for further optimizing PDMS for hES cell culture, moving towards the use of microdevices in establishing precise control over stem cell fate. The second study explored the use of an easily constructed diffusion-based device to grow hES cells in culture on a defined, physiologic oxygen (O₂) gradient. We observed greater hES cell survival and higher levels of pluripotency markers in the lower O₂ regions of the gradient. The greatest benefit was observed at O₂ levels below 5%, narrowing the potential optimal range of O₂ for the maintenance of pluripotent hES cells. Finally, we developed a small molecule-mediated adherent and feeder-free neural differentiation protocol that reduced the cost and time scale for in vitro differentiation of neural precursors and functional neurons from human pluripotent cells. hES cell-derived neural precursors transplanted into a murine model of focal ischemic stroke survived, improved neurogenesis, and differentiated into neurons. Transplant also led to a more consistent and measurable sensory recovery after stroke as compared to untransplanted controls. This protocol represents a potentially translatable method for the generation of CNS progenitors from human pluripotent stem cells.
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Beligala, Dilshan Harshajith. "Stem-like cells and glial progenitors in the adult mouse suprachiasmatic nucleus". Bowling Green State University / OhioLINK, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1566319291491512.
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Okolicsanyi, Rachel K. "Mesenchymal stem cells as mediators of the neuronal cell niche". Thesis, Queensland University of Technology, 2015. https://eprints.qut.edu.au/84485/1/Rachel_Okolicsanyi_Thesis.pdf.
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This study examined the role of heparan sulfate proteoglycans (HSPGs) in neural lineage differentiation of human mesenchymal stem cells (hMSCs). Several HSPGs were identified as potential new targets controlling neural fate specification and may be applied to the development of improved models to examine and repair brain damage. hMSCs were characterised throughout extended in vitro expansion for neural lineage potential (neurons, astrocytes, oligodendrocytes) and differentiated using terminal differentiation and intermediate sphere formation. Brain damage and neurological disorders caused by injury or disease affect a large number of people often resulting in lifelong disabilities. Multipotent mesenchymal stem cells have a large capacity for self-renewal and provide an excellent model to examine the regulation and contribution of both stem cells and their surrounding microenvironment to the repair of neural tissue damage.
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Tasneem, Sameera. "EFFECTS OF ENVIRONMENTAL HEAVY METALS ON NERUAL STEM CELL SURVIVAL AND DIFFERENTIATION". Cleveland State University / OhioLINK, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=csu1400500632.
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Steeg, Rachel. "The role of low oxygen in the self-renewal and neural differentiation of human pluripotent stem cells". Thesis, Cardiff University, 2015. http://orca.cf.ac.uk/69784/.
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Human embryonic stem cells are derived from the pre-implantation blastocyst, residing in an extremely low oxygen environment. Application of this in vivo oxygen concentration to an in vitro setting has previously shown to be essential for driving the self-renewal and directed differentiation of human pluripotent stem cells. However, studies on hESCs at low oxygen have been frequently contra-indicatory. Here, comparative analysis of a hESC and iPSC line at a low oxygen concentration resulted in divergent effects across the two cell lines. Augmented TGFβ signalling was observed in conjunction with up-regulated transcription of pluripotency markers NANOG and POU5F1 in hESCs but not iPSCs. IPSCs also shifted to a state of increased proliferation whereas hESCs did not. It was also observed that exposure of hESCs and iPSCs to light throughout low oxygen culture induced large amounts of apoptosis, highlighting the requirement for careful selection of cell culture equipment for environmental oxygen control. Both the embryonic and adult brain retain tissue specific oxygen concentrations far below that at atmospheric oxygen concentrations. Previous studies showed that embryonic silencing of factors responsive to low oxygen caused a range of embryonic abnormalities, including deformation of the neural plate and tube. Here, differentiation of hESCs to an early neuroectodermal identity at low O2 did not definitively augment production of NPCs but did additively suppress BMP signalling above that at atmospheric O2. A concordant rise in apoptosis was also observed in a manner both independent of, and augmented by, exogenous BMP inhibition. Subsequently, neurogenesis at low oxygen produced terminal neurons with accelerated and augmented synaptic and induced excitability. This was characterised by a rise in the rate of membrane depolarisation, increased action potential overshoot and accelerated expression of pre-synaptic marker Synaptophysin. These results highlight a novel, critical role for low oxygen in augmenting the excitability of hESC derived neurons.
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McLaughlin, Heather Ward. "Modeling sporadic Alzheimer's disease using induced pluripotent stem cells". Thesis, Harvard University, 2014. http://nrs.harvard.edu/urn-3:HUL.InstRepos:13094355.
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Despite being the leading cause of neurodegeneration and dementia in the aging brain, the cause of Alzheimer's disease (AD) remains unknown in most patients. The terminal pathological hallmarks of abnormal protein aggregation and neuronal cell death are well-known from the post-mortem brain tissue of Alzheimer's disease patients, but research into the earliest stages of disease development is hindered by limited model systems. In this thesis, an in vitro human neuronal system was derived from induced pluripotent stem (iPS) cell lines reprogrammed from dermal fibroblasts of AD patients and age-matched controls. This allows us to investigate the cellular mechanisms of AD neurodegeneration in the human neurons of sporadic AD (SAD) patients, whose development of the disease cannot be explained by our current understanding of AD. We show that neural progenitors and neurons derived from SAD patients show an unexpected expression profile of enhanced neuronal gene expression resulting in premature differentiation in the SAD neuronal cells. This difference is accompanied by the decreased binding of the repressor element 1-silencing transcription/neuron-restrictive silencer factor (REST/NRSF) transcriptional inhibitor of neuronal differentiation in the SAD neuronal cells. The SAD neuronal cells also have increased production of \(amyloid-\beta\) and higher levels of tau protein, the main components of the plaques and tangles in the AD brain.
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Eiriz, Maria Francisca Santos. "Migration and differentiation of neuronal precursors in the postnatal brain: insights from the subventricular zone and cerebellum". Doctoral thesis, Faculdade de Ciências e Tecnologia, 2013. http://hdl.handle.net/10362/11447.
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Fundação para a Ciência e a Tecnologia - SFRH/BD/42848/2008, através do Programa MIT_Portugal em Sistemas de Bioengenharia; projectos PTDC/SAUNEU/104415/2008 e Projecto ref. 96542 da Fundação Caloust Gulbenkian
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Bertacchi, Michele. "In vitro neural differentiation of mouse embryonic stem cells: the positional identity of mouse ES-generated neurons is affected by BMP, Wnt and activin signaling". Doctoral thesis, Scuola Normale Superiore, 2014. http://hdl.handle.net/11384/85976.
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Carradori, Dario. "Novel nanoparticle-based drug delivery system for neural stem cell targeting and differentiation". Thesis, Angers, 2017. http://www.theses.fr/2017ANGE0056/document.
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Les cellules souches neurales (CSNs) se situent dans des régions spécifiques du système nerveux central qui sont appelées niches. Ces cellules sont capables de se répliquer ou se différentier en cellules neurales spécialisées (neurones, astrocytes et oligodendrocytes). C’est grâce à cette propriété de différentiation que les CSNs sont étudiées comme thérapie chez les patients atteints d’une maladie neurodégénérative. En effet, elles pourraient remplacer les cellules neurales altérées et ainsi restaurer les fonctions neurologiques. De nombreuses approches ont été développées afin de stimuler la différentiation des CSNs, dont la plus prometteuse est la différentiation des cellules endogènes directement au sein de leurs niches. Actuellement, il n’existe pas de molécule active ou de système thérapeutique qui cible les CSNs endogènes et qui induit leur différentiation simultanément. Le but de ce travail est de fournir un système de délivrance de molécules bioactives capable de cibler les CSNs endogènes et d'induire leur différenciation in situ. Nous avons développé et caractérisé des nanoparticules lipidiques (LNC), un système de délivrance très versatile. NFL-TBS.40-63, un peptide ciblant les CSNs, a été adsorbé à la surface des LNC afin de les diriger contre les CSNs endogènes. Nous avons observé que ces NFL-LNC ne ciblaient que les CSNs du cerveau et pas de la moelle. Afin d’étudier les interactions spécifiques entre les nanoparticules et les CSNs, nous avons caractérisé et comparé les propriétés de leur membrane plasmique. Enfin, nous avons encapsulé de l’acide rétinoïque, une molécule connue pour stimuler la différentiation des CSNs, dans les LNC-NFL et étudié leur impact sur la différentiation de CSNs in vitro et in vivo. Ce travail contribue au développement de thérapies efficaces et sures pour le traitement de maladies neurodégénératives à travers la différentiation de CSNs endogènesNeural stem cells (NSCs) are located in specific regions of the central nervous system called niches. Those cells are able to self-renew and to differentiate into specialized neuronal cells (neurons, astrocytes and oligodendrocytes). Due to this differentiation property, NSCs are studied to replace neuronal cells and restore neurological functions in patients affected by neurodegenerative diseases. Several therapeutic approaches have been developed and endogenous NSC stimulation is one of the most promising. Currently, there is no active molecule or therapeutic system targeting endogenous CSNs and inducing their differentiation at the same time. The aim of the work was to provide a drug delivery system able both to target endogenous CSNs and to induce their differentiation in situ. Here, we developed and characterized lipidic nanoparticles (LNC) targeting endogenous NSCs. A peptide called NFL-TBS.40-63, known for its affinity towards NSCs, was adsorbed at the surface of LNC. We observed that NFL-LNC specifically targeted NSC from the brain and not from the spinal cord in vitro and in vivo. To explain this specificity, we characterized and compared NFL-LNC interactions with the plasmatic membrane of both cell types. Finally, we demonstrated that by loading retinoic acid in NFL-LNC we were able to induce brain NSC differentiation in vitro and in vivo. This work contributes to the development of efficient and safe therapies for the treatment of neurodegenerative disease via the differentiation of endogenous NSCs