Dissertations / Theses on the topic 'Neural cultures'
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De, La Garza Richard. "Determination of neuronal morphology in spinal monolayer cultures." Thesis, University of North Texas, 1989. https://digital.library.unt.edu/ark:/67531/metadc798395/.
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The objective of the completed research was to characterize the morphology of individual neurons within monolayer networks of fetal mouse spinal tissue via intraperikaryal injections of horseradish peroxidase (HRP). Thirty labelled neurons were reconstructed via camera lucida drawings and morphometrically analyzed.
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Amaral, Ana Isabel Porém. "Metabolic flux analysis of neural cell metabolism in primary cultures." Doctoral thesis, Universidade Nova de Lisboa. Instituto de Tecnologia Química e Biológica, 2011. http://hdl.handle.net/10362/6849.
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Dissertation presented to obtain the Ph.D degree in Biochemistry, NeuroscienceBrain energy metabolism results from a complex group of pathways and trafficking mechanisms between all cellular components in the brain, and importantly provides the energy for sustaining most brain functions. In recent decades, 13C nuclear magnetic resonance (NMR) spectroscopy and metabolic modelling tools allowed quantifying the main cerebral metabolic fluxes in vitro and in vivo. These investigations contributed significantly to elucidate neuro-glial metabolic interactions, cerebral metabolic compartmentation and the individual contribution of neurons and astrocytes to brain energetics. However, many issues in this field remain unclear and/or under debate.
To the financial support provided by Fundação para a Ciência a Tecnologia (SFRH/BD/29666/2006; PTDC/BIO/69407/2006) and to the Clinigene – NoE (LSHBCT2006- 010933). I further acknowledge the Norwegian Research Council for a fellowship that allowed me to perform part of my PhD work at NTNU, Norway.
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Cullen, Daniel Kacy. "Traumatically-Induced Degeneration and Reactive Astrogliosis in 3-D Neural Co-Cultures: Factors Influencing Neural Stem Cell Survival and Integration." Diss., Georgia Institute of Technology, 2005. http://hdl.handle.net/1853/7584.
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Traumatic brain injury (TBI) results from a physical insult to the head and often results in temporary or permanent brain dysfunction. However, the cellular pathology remains poorly understood and there are currently no clinically effective treatments. The overall goal of this work was to develop and characterize a novel three-dimensional (3-D) in vitro paradigm of neural trauma integrating a robust 3-D neural co-culture system and a well-defined biomechanical input representative of clinical TBI. Specifically, a novel 3-D neuronal-astrocytic co-culture system was characterized, establishing parameters resulting in the growth and vitality of mature 3-D networks, potentially providing enhanced physiological relevance and providing an experimental platform for the mechanistic study of neurobiological phenomena. Furthermore, an electromechanical device was developed that is capable of subjecting 3-D cell-containing matrices to a defined mechanical insult, with a predicted strain manifestation at the cellular level. Following independent development and validation, these novel 3-D neural cell and mechanical trauma paradigms were used in combination to develop a mechanically-induced model of neural degeneration and reactive astrogliosis. This in vitro surrogate model of neural degeneration and reactive astrogliosis was then exploited to assess factors influencing neural stem cell (NSC) survival and integration upon delivery to this environment, revealing that specific factors in an injured environment were detrimental to NSC survival. This work has developed enabling technologies for the in vitro study of neurobiological phenomena and responses to injury, and may aid in elucidating the complex biochemical cascades that occur after a traumatic insult. Furthermore, the novel paradigm developed here may provide a powerful experimental framework for improving treatment strategies following neural trauma, and therefore serve as a valid pre-animal test-bed.
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Cullen, Daniel Kacy. "Traumatically-induced degeneration and reactive astrogliosis in three-dimensional neural co-cultures." Available online, Georgia Institute of Technology, 2005, 2005. http://etd.gatech.edu/theses/available/etd-11282005-210117/.
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Thesis (Ph. D.)--Biomedical Engineering, Georgia Institute of Technology, 2006.Robert McKeon, Committee Member ; Robert Lee, Committee Member ; Robert Guldberg, Committee Member ; Ravi Bellamkonda, Committee Member ; Michelle LaPlaca, Committee Chair. Vita.
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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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Rioult-Pedotti, Marc Guy. "Optical multisite recording of neural activity patterns in organotypic spinal cord tissue cultures /." [S.l.] : [s.n.], 1991. http://e-collection.ethbib.ethz.ch/show?type=diss&nr=9393.
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Olivero, Daniel. "Traumatic brain injury biomarker discovery using mass spectrometry imaging of 3D neural cultures." Thesis, Georgia Institute of Technology, 2011. http://hdl.handle.net/1853/41102.
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Biomarker research is of great interest in the field of traumatic brain injury (TBI), since there are numerous potential markers that may indicate central nervous system damage, yet the brain is normally well isolated and discovery is at its infancy. Traditional methods for biomarker discovery include time consuming multi step chromatographic mass spectrometery (MS) techniques or pre-defined serial probing using traditional assays, making the identification of biomarker panels limiting and expensive. These shortfalls have motivated the development of a MS based probe that can be embedded into 3D neural cultures and obtain temporal and spatial information about the release of biomarkers. Using the high sensitivity MS ionization method of nano-electrospray ionization (nano-ESI) with an in-line microdialysis (MD) unit allows us to use MS to analyze low concentrations of TBI biomarkers from within cell cultures with no need for off-line sample manipulation. This thesis goes through the development of the probe by studying the theoretical principles, simulations and experimental results of the probe's capability to sample small local concentrations of a marker within cell culture matrix, the MD unit's sample manipulation capabilities, and the ability to detect markers using in-line MD-nano-ESI MS.
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Bakkum, Douglas James. "Dynamics of embodied dissociated cortical cultures for the control of hybrid biological robots." Diss., Atlanta, Ga. : Georgia Institute of Technology, 2007. http://hdl.handle.net/1853/22596.
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Thesis (Ph. D.)--Mechanical Engineering, Georgia Institute of Technology, 2008.Committee Chair: Steve M. Potter; Committee Member: Eric Schumacher; Committee Member: Robert J. Butera; Committee Member: Stephan P. DeWeerth; Committee Member: Thomas D. DeMarse.
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Madhavan, Radhika. "Role of spontaneous bursts in functional plasticity and spatiotemporal dynamics of dissociated cortical cultures." Diss., Atlanta, Ga. : Georgia Institute of Technology, 2007. http://hdl.handle.net/1853/24756.
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Thesis (Ph.D.)--Biomedical Engineering, Georgia Institute of Technology, 2007.Committee Chair: Potter, Steve; Committee Member: Butera, Robert; Committee Member: DeWeerth, Stephen; Committee Member: Schumacher, Eric; Committee Member: Wenner, Pete.
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McLoughlin, Justin. "A novel in vitro shear device for inducing high strain rate deformation on neural cell cultures." Thesis, Georgia Institute of Technology, 2000. http://hdl.handle.net/1853/16011.
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Park, Deric M., Jinkyu Jung, Jimmy Masjkur, Stylianos Makrogkikas, Doreen Ebermann, Sarama Saha, Roberta Rogliano, et al. "Hes3 regulates cell number in cultures from glioblastoma multiforme with stem cell characteristics." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2013. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-127014.
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Tumors exhibit complex organization and contain a variety of cell populations. The realization that the regenerative properties of a tumor may be largely confined to a cell subpopulation (cancer stem cell) is driving a new era of anti-cancer research. Cancer stem cells from Glioblastoma Multiforme tumors express markers that are also expressed in non-cancerous neural stem cells, including nestin and Sox2. We previously showed that the transcription factor Hes3 is a marker of neural stem cells, and that its expression is inhibited by JAK activity. Here we show that Hes3 is also expressed in cultures from glioblastoma multiforme which express neural stem cell markers, can differentiate into neurons and glia, and can recapitulate the tumor of origin when transplanted into immunocompromised mice. Similar to observations in neural stem cells, JAK inhibits Hes3 expression. Hes3 RNA interference reduces the number of cultured glioblastoma cells suggesting a novel therapeutic strategy.
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Teller, Amado Sara. "Functional organization and networ resilience in self-organizing clustered neuronal cultures." Doctoral thesis, Universitat de Barcelona, 2016. http://hdl.handle.net/10803/396114.
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Major dynamical traits of a neuronal network are shaped by its underlying circuitry. In several neurological disorders, the deterioration of brain's functionality and cognition has been ascribed to changes in the topological properties of the brain's circuits. To deepen in the understanding of the activity-connectivity relationship, neuronal cultures have emerged as remarkable systems given their accessibility and easy manipulation. A particularly appealing configuration of these in vitro systems consists in an assembly of interconnected aggregates of neurons termed 'clustered neuronal networks'.
These networks exhibit a complex dynamics in which clusters fire in small groups, shaping communities with rich spatiotemporal properties. The detailed characterization of this dynamics, as well as its resilience to perturbations, has been the main objective of this thesis. In our experiments we monitored spontaneous activity using calcium fluorescence imaging, which allows the detection of neuronal firing events with both high temporal and spatial resolution. The detailed analysis of the recorded activity, in the context of network theory and community analysis, allowed for the quantification of important properties, including the effective connectivity map and its major topological descriptors.
As major results, we observed that these clustered networks present hierarchical modularity, assortative mixing and the presence of a rich club core, a series of features that have also been observed at the scale of the brain. All these characteristic topological traits are associated with a robust architecture that reinforces and stabilizes network activity. To verify the existence of such robustness in our cultures, we studied their resilience upon chemical and physical damage. We concluded that, indeed, clustered networks present higher resilience compared to other configurations. Moreover, these clustered networks exhibited recovery mechanisms that can be linked to the balance between integration and segregation in the network, which ultimately tend to preserve network activity upon damage. Thus, these in vitro preparations offer a unique scenario to explore vulnerability in networks with topological properties similar to the brain. Moreover, the combination of all these approaches can help to develop models to quantify damage upon network degradation, with promising applications for the study of neurological disorders in vitro.Desvelar la relación entre la red de conexiones anatómica y su emergente dinámica es uno de los grandes desafíos de la neurociencia actual. En este sentido, los cultivos neuronales han tomado un papel muy importante para entender esta cuestión, ya que fenomenologías fundamentales pueden ser estudiadas a escalas más tratables. Los cultivos neuronales se obtienen típicamente a base de disociar tejido neuronal de una parte específica del cerebro, corteza cerebral de rata en nuestro caso, y su cultivo en un medio adecuado. Neuronas en cultivo constituyen en 1-2 semanas una red nueva con una actividad espontánea rica. Una de las preparaciones in vitro que ofrece mayor potencial es las 'redes clusterizadas'. Estas redes se auto-organizan de forma natural, formando grupos de neuronas (clústeres) interconectados a través de axones. La caracterización de la dinámica de estas redes clusterizadas, así como su sensibilidad a perturbaciones, ha sido el objetivo principal de esta tesis. Así, hemos caracterizado la red funcional del cultivo a partir de su dinámica espontánea, desarrollando para ello un novedoso modelo fisicomatemático. Hemos observado que las redes tienen una conectividad modular, donde clústeres tienden a conectarse fuertemente en pequeños grupos, los cuales a su vez se conectan entre ellos. Además, las redes funcionales muestran propiedades topológicas clave, en especial asortatividad (interconexión preferente de clústeres con número similar de conexiones) y la existencia de un 'rich club' (grupo de clústeres con una interconectividad tan destacada que forman el núcleo fundamental de la red). Estas propiedades confieren una gran robustez y flexibilidad a la red. Por esta razón, en la tesis hemos investigado diferentes perturbaciones físicas y bioquímicas, demostrando que las redes clusterizadas son mucho más resistentes a daño que otras configuraciones, lo que refuerza la relación entre las propiedades topológicas descritas y resistencia al daño. Además, observamos que las redes presentaron diferentes mecanismos de reforzamiento entre conexiones para preservar la actividad de la red. Por ello, las redes clusterizadas constituyen una plataforma ideal para estudiar resistencia en redes o como sistema modelo aplicado a estudios de enfermedades neurodegenerativas, como por ejemplo Alzheimer.
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Sparks, Christopher A. "Investigations of neuronal network responses to electrical stimulation in murine spinal cultures." Thesis, University of North Texas, 2001. https://digital.library.unt.edu/ark:/67531/metadc3027/.
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Spontaneous activity in neuronal networks in vitro is common and has been well documented. However, alteration of spontaneous activity in such networks via conditioning electrical stimulation has received much less experimental attention. Two different patterns of electrical stimulation were used to enhance or depress the level of spontaneous activity in spinal cord cultures. High-frequency stimulation (HFS), a method routinely shown to increase the efficacy of synaptic transmission, was employed to augment spontaneous activity. Low-frequency stimulation (LFS), the technique often applied to depress synaptic efficacy, was employed to decrease spontaneous activity. In addition, LFS was used to reverse the effect of HFS on spontaneous activity. Likewise, HFS was applied to counter the effect of LFS. Because these networks were grown on multi-microelectrode plates (MMEPs), this allowed the simultaneous stimulation of any combination of the 64 electrodes in the array. Thus, the possible differences in response to single versus multi-electrode stimulation were also addressed. Finally, test-pulses were delivered before and after the conditioning stimulation on the same stimulation electrode(s) in order to assess the change in mean evoked action potentials (MEAPs). Dissociated spinal tissue from embryonic mice was allowed to mature into self-organized networks that exhibited spontaneous bursting activity after two weeks of incubation. Spontaneous activity was monitored from up to 14 recording channels simultaneously. Although uniform responses to stimulation across all recording electrodes were rarely observed, a large majority of the recording channels had similar responses. Spontaneous activity was increased in 52% of 89 HFS trials, whereas activity was decreased in 35% of 75 LFS trials. The duration of most of these increases was less than 5 minutes. When there were substantial and long-term (> 15 min) changes in spontaneous activity, the opposing stimulation pattern successfully reversed the effect of the previous stimulation. The percent change in MEAPs following conditioning stimulation suggested that synaptic modification had taken place in 75% of all test-pulse stimulation trials.
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Labour, Marie-Noëlle. "Collagène auto-assemblé en support 3D biomimétique fonctionnalisé pour la différenciation de cellules nerveuses." Thesis, Montpellier 1, 2012. http://www.theses.fr/2012MON13508/document.
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L'objectif de ce travail était de mettre au point un système de culture tridimensionnel compartimenté pour la différentiation de cellules neurales et la croissance des neurites en 3D. Les matériaux biomimétiques permettent l'élaboration de microenvironnements contrôlés qui peuvent orienter la réponse cellulaire. Ils sont particulièrement intéressants pour les études fondamentales visant à étudier des voies de signalisation impliquées dans des processus physiologiques ou pathologiques. Nous nous sommes intéressés à la maladie d'Alzheimer, où l'on observe des neurites dystrophiques associés aux plaques amyloïdes. Aucune relation n'a été réellement établie entre l'interaction neurites - agrégats, leur dystrophie et la mort neuronale. Dans un premier temps, nous avons décrit et caractérisé la structure et les propriétés de matrices de collagène fibrillaire d'épaisseur calibrée. Ensuite, nous avons mis au point la fonctionnalisation de ces matrices avec des facteurs de croissance neurotrophiques (NGF et BDNF). Deux techniques ont été étudiées : l'imprégnation/libération et le couplage covalent. Ces matrices fonctionnalisées ont été validées comme support pour la différenciation de cellules nerveuses (PC-12 et SH-SY5Y) par des études de la morphologie cellulaire. Enfin, nous avons caractérisé des agrégats amyloïdes (Aβ) formés à l'intérieur des matrices de collagène par co-précipitation du peptide Aβ avec le collagène et nous avons étudié leur toxicité sur les cellules neuralesThe objective of this work was to develop a 3D compartmented cell culture set-up that allow the differentiation of nerve cells and the growth of neurites in the matrix depth. Biomimetic materials enable the formation of controlled microenvironments that orient cell behavior. They are particularly interesting for fundamental studies that aim to study signaling pathways involved in physiologic or pathologic processes. We focused on Alzheimer's disease, in which dystrophic neurites are associated to amyloid plaques. No direct relationship has yet been established between Aβ aggregates-neurite interaction, neurite dystrophy and cell death. First, we described and characterized the structure and properties of fibrillar collagen matrices with adapted thickness. Then, we adjusted functionalization of these matrices with neurotrophic growth factors (NGF and BDNF). Two methods were studied: impregnation/release and covalent coupling. Cell morphology studies confirmed that these functionalized matrices were efficient supports for nerve cells differentiation (PC-12 and SH-SY5Y). Finally, we have characterized Aβ aggregates that were formed inside collagen matrices by coprecipitation of amyloid peptide and collagen and we studied their toxicity on neural cells
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Luca, Luminita Eugenia. "Oxygen Glucose Deprivation and Hyperthermia Induce Cellular Damage in Neural Precursor Cells and Immature Neurons." Scholarly Repository, 2008. http://scholarlyrepository.miami.edu/oa_dissertations/184.
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Hyperthermia damages both developing and adult brains, especially when it occurs after ischemia or stroke. Work presented in this dissertation used in vitro models of these stresses to investigate mechanisms underlying damage to immature neurons and neural precursors cultured from embryonic rat brain. Studies described in Chapter 2 investigated the effects of a brief, intense hyperthermic stress (30-45 min at 43ºC). This stress produced a selective depletion of nestin-immunoreactive neural precursor cells, and reduced proliferation, as evidenced by reduced BrdU incorporation into young Tuj1-immunoreactive neurons. The stress activated caspase 3, and produced multiple signs of nuclear damage as well as early and persisting mitochondrial depolarization. Cycloheximide, an inhibitor of protein synthesis, reduced cell death. All these findings suggest an apoptotic death process. Studies described in Chapter 3 used a combination of oxygen-glucose deprivation (OGD, 2 h) followed by mild 41ºC hyperthermia for 90 min (T). The combined OGDT stress reduced both survival in monolayer cultures and colony-forming ability in neurospheres. Cell death occurred gradually over 2 days, and was accompanied by caspase activation that began within 6 h post-stress. Post-stress application of cycloheximide or a general caspase inhibitor (especially qVD-OPH) reduced cell death, but specific inhibitors of caspases 2, 3, 8 or 9 were ineffective. OGDT led to upregulation of the pro-apoptotic protein Bim as well as redistribution of Bax from cytoplasm to mitochondria within 6 h. Persisting mitochondrial depolarization began within 3 h following the combined OGDT stress, but not following individual OGD or T stresses alone. These findings suggest that OGD sensitizes neural precursor cells to hyperthermia-induced damage, and that the combined OGDT stress kills neural precursors via apoptotic mechanisms that include activation of mitochondrial death pathways. Results of these studies suggest that immature neurons and neural precursors are especially vulnerable to hyperthermia-induced damage via apoptotic mechanisms. Pan-caspase inhibitors may be a promising therapeutic strategy to preserve viability of these cells following stroke with hyperthermia.
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Vernekar, Varadraj Nagesh. "Optimization of 3-d neural culture and extracellular electrophysiology for studying injury-induced morphological and functional changes." Diss., Georgia Institute of Technology, 2010. http://hdl.handle.net/1853/39624.
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This work characterized an in vitro 3-D neural co-culture model electrophysiologically via multi electrode arrays (MEAs), and morphologically via immunocytochemistry. Since MEA surface insulation SU-8 2000 can be used in neural micro- and multi- electrode arrays, this investigation first developed techniques to make SU-8 2000 cytocompatible. The in vitro 3-D neural co-culture model was then used to study viability and electrophysiological responses to physical injury as well as drugs known to affect network signaling. 1) SU-8 2000 cytotoxicity to neuronal cultures was linked to both poor adhesive properties and toxic components, such as solvents and photo acid generator elements. Surface treatments of oxygen plasma or parylene coating following optimal combinations of heat and isopropanol sonication showed improvement in SU-8 2000 cytocompatibility. 2) The 3-D neural networks within the 3-D co-cultures maintained considerable process outgrowth and complex 3-D structure. The cultures were viable up to three weeks in vitro with functional synaptic connections and spontaneous electrophysiological activity that was responsive to chemical modulation. This electrophysiological activity was modulated by synaptic inhibition. 3) Injury experiments demonstrated that both shear and compression loading significantly increased acute membrane permeability of cells in a strain rate dependent manner. Cell death correlated with higher membrane permeability, and shear resulted in more death than compression in these 3-D cultures. While techniques were developed for making a major micro-fabrication material cytocompatible, engineering the 3-D neural co-culture resulted in a more physiologically-representative neural tissue platform, allowing an increased understanding of structure-function relationships. Overall, this research established and characterized a neural culture system for the mechanistic study of cell growth, cell-cell and cell-matrix interactions, as well as the responses to chemical or mechanical perturbations. This is the first investigation of the network-level electrophysiological activity of 3-D dissociated cultures. This system can be used to model various pathological states in vitro, testing various reparative drugs; cell-, and tissue-engineering based strategies; as well as for pre-animal and pre-clinical testing of neural implants.
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Rambani, Komal. "Thick brain slice cultures and a custom-fabricated multiphoton imaging system: progress towards development of a 3D hybrot model." Thesis, Georgia Institute of Technology, 2007. http://hdl.handle.net/1853/22702.
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Development of a three dimensional (3D) HYBROT model with targeted in vivo like intact cellular circuitry in thick brain slices for multi-site stimulation and recording will provide a useful in vitro model to study neuronal dynamics at network level. In order to make this in vitro model feasible, we need to develop several associated technologies. These technologies include development of a thick organotypic brain slice culturing method, a three dimensional (3D) micro-fluidic multielectrode Neural Interface system (µNIS) and the associated electronic interfaces for stimulation and recording of/from tissue, development of targeted stimulation patterns for closed-loop interaction with a robotic body, and a deep-tissue non-invasive imaging system. To make progress towards this goal, I undertook two projects: (i) to develop a method to culture thick organotypic brain slices, and (ii) construct a multiphoton imaging system that allows long-term and deep-tissue imaging of two dimensional and three dimensional cultures.
Organotypic brain slices preserve cytoarchitecture of the brain. Therefore, they make more a realistic reduced model for various network level investigations. However, current culturing methods are not successful for culturing thick brain slices due to limited supply of nutrients and oxygen to inner layers of the culture. We developed a forced-convection based perfusion method to culture viable 700µm thick brain slices.
Multiphoton microscopy is ideal for imaging living 2D or 3D cultures at submicron resolution. We successfully fabricated a custom-designed high efficiency multiphoton microscope that has the desired flexibility to perform experiments using multiple technologies simultaneously. This microscope was used successfully for 3D and time-lapse imaging.
Together these projects have contributed towards the progress of development of a 3D HYBROT.
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3D Hybrot: A hybrid system of a brain slice culture embodied with a robotic body.
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Garrido-Sanchez, Luis Edmundo. "Conception et mise en œuvre d'un système d'aide a la modélisation de procédés de fermentation utilisant un système expert et des réseaux de neurones." Vandoeuvre-les-Nancy, INPL, 1993. http://www.theses.fr/1993INPL023N.
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L'objectif général de notre travail a été de développer une méthode de structuration des connaissances procédurales mises en œuvre lors de la modélisation de bioprocédés. Ce modèle cognitif permet la réalisation d'un système informatique d'aide à la modélisation. Dans une première partie, nous développons la méthodologie suivie pour effectuer l'extraction du savoir faire d'un expert en modélisation. Nous présentons l'expertise obtenue ainsi qu'une description de la procédure de modélisation et les éléments cinétiques utilisés. Dans une deuxième partie nous proposons la structure d'un système hybride compose d'un système expert contenant le savoir faire en construction de modèles et de plusieurs réseaux de neurones intégrant les connaissances nécessaires a la classification des courbes cinétiques. La troisième partie traite du test des procédures du système dans le cas d'une culture de bactéries pour la production d'acide lactique. Un modèle cinétique de la littérature est utilise pour simuler les données fournies au système: le modèle restitue lui est alors comparé. Finalement, nous présentons le modèle obtenu avec le prototype du système d'aide à la modélisation a partir de données expérimentales obtenues dans le cas d'une culture continue de kluyveromyces fragilis
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Morefield, Samantha I. (Samantha Irene). "Responses of Cultured Neuronal Networks to the Cannabinoid Mimetic Anandamide." Thesis, University of North Texas, 1998. https://digital.library.unt.edu/ark:/67531/metadc277717/.
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The effects of cannabinoid agonists on spontaneous neuronal network activity were characterized in murine spinal cord and auditory cortical cultures with multichannel extracellular recording using photoetched electrode arrays. Different cultures responded reproducibly with global decreases of spiking and bursting to anandamide and methanandamide, but each agonist showed unique minor effects on network activity. The two tissues responded in a tissue-specific manner. Spontaneous activity in spinal tissue was terminated by 1 μM anandamide and 6.1 μM methanandamide. Cortical activity ceased at 3.5 μM and 2.8 μM respectively. Irreversible cessation of activity was observed beyond 8 μM for both tissues and test substances. Palmitoylethanolamide, demonstrated that CB2 receptors were not present or not responsive. However, the data strongly suggested the presence of CB1 receptors.
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Beattie, Allison Jane. "Organised neural networks in culture." Thesis, University of Glasgow, 2006. http://theses.gla.ac.uk/1921/.
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The aim of my research was to recreate simple, spatially organised neural networks in culture for the study of neural network behaviour. Spinal cord neurons were chosen as the biological model, as much is already known about spinal cord tissue circuitry in-vivo. These simple networks of cells were created by chemical patterning techniques (micro-contact printing (mCP)), and topographical guidance mechanisms. mCP was used to test the hypothesis that alterations in network architecture could affect network behaviour. Changes in network structures were identified, using immunocytochemical staining and scanning electron microscopy (SEM). Of the six patterns tested it was concluded that the Jude pattern did not satisfy the criteria required for a neural network. Cells failed to comply to the extreme angles of this design and so a hexagonal pattern was introduced. Dendritic architecture, of varying designs, was incorporated into these hexagonal networks with the aim of determining if variation in dendritic arborisation could affect network activity. An analysis of the result showed that cell morphology and connectivity was visibly altered, suggesting network characteristics were affected. An attempt was made to create organised nerve cultures using micro-metric grooved patterns in poly-dimethylsiloxane (PDMS). The cellular response was determined by immunocytochemical staining and SEM imaging. Cells grown on micrometric topographical patterns did not align within the grooves as predicted. Therefore the effect of nano-metric pillared topography, created in poly-caprolactone, on nerve cell guidance was investigated. In comparison to the flat material, this nanotopography reduced cell adhesion, although it was not completely non-adhesive. After 1 week cells were visibly aligning to the topography, at the micro-and nanometric level, and appeared to be growing longer processes compared to the cells grown on flat structures. This result suggests nanopillared topography has a promising future in nerve guidance studies.
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Murphy, Eric James. "Cell culture models for neural trauma /." The Ohio State University, 1989. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487672245901403.
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Ishikawa, Masaaki. "Transplantation of neurons derived from human iPS cells cultured on collagen matrix into guinea-pig cochleae." 京都大学 (Kyoto University), 2017. http://hdl.handle.net/2433/225472.
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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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Guo, Aobo. "Progranulin (PGRN) functions in neuronal cultures." Thesis, University of British Columbia, 2009. http://hdl.handle.net/2429/12702.
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Null mutations in the progranulin gene (PGRN) have been identified as a major cause of frontotemporal lobe dementia with ubiquitinated inclusions (FTLD-U). In this disorder, the ubiquitinated aggregated protein inclusions of a normally nuclear-located RNA processing protein called TAR DNA binding protein (TDP-43) accumulate in the cytoplasm of neurons. To determine whether aspects of this clinical pathology can be established in primary cultures of mouse cortical neurons, PGRN levels were knocked down in cultures using lentiviral vectors to introduce siRNA constructs. Similar to observations in the brains of FTLD-U patients, TDP-43 levels were markedly decreased in the nucleus of PGRN knockdown neurons relative to cytoplasmic levels in comparison to control neurons and significantly recovered by over-expressed PGRN, although the TDP-43 cleavage was not obvious in this model. Meanwhile, depletion of PGRN elevated levels of activated caspase-3, and the PGRN-deficient neurons demonstrated enhanced vulnerability to sub-lethal doses of NMDA and H₂O₂. These results show that it is possible to recapitulate key features of the PGRN null mutation in a culture dish within a short period of time and suggest that the seeds of this form of frontotemporal dementia may be sown early in life.
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Eriksson, Malin. "Manipulating neural stem cells." Stockholm, 2010. http://diss.kib.ki.se/2010/978-91-7409-853-2/.
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Corradini, Daniele. "Statistical characterization of cultured neural networks activity recorded via MEA." Master's thesis, Alma Mater Studiorum - Università di Bologna, 2020. http://amslaurea.unibo.it/20380/.
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In questa tesi si sono analizzate le attività di neuroni in coltura accoppiati con un Multi-Electrode Array (MEA) a 60 canali. Le reti neurali in vitro sono influenzate da diversi fattori, come la densità di coltura, l'età e l'animale di provenienza dei neuroni.
Lo scopo di questa tesi è quello di caratterizzare questa variabilità attraverso varie analisi statistiche.
Utilizzando un software sviluppato in Python si sono estratte 40 misure per descrivere un ampio spettro delle attività delle cellule, come lo spiking e i network bursts.
Sono state caratterizzate le distribuzioni statistiche di queste feature, ed una analisi PCA per vedere la segregazione dei dati nelle classi considerate.
Sono poi state implementate una analisi con K-Means Clustering e una classificazione mediante Random Forest, per caratterizzare la separazione spontanea e mediante label dei vari tipi di misurazioni.
É stato studiato anche l'effetto del sottocampionamento temporale sulle misure, determinando che anche con un quarto della serie temporale disponibile si sono potute estrarre misure significative. Un altro tipo di sottocampionamento preso in considerazione è stato quello spaziale. Anche qui le misure si sono dimostrate robuste riducendo il numero di elettrodi da cui estrarre il segnale, fino ad un minimo di 10 elettrodi.
Questa analisi della robustezza alla variabilità e al sottocampionamento delle misure di attività spontanea di reti di neuroni in vitro mediante setup MEA é utile per studiare gli effetti di sostanze chimiche, stimoli elettrici o malattie sulle colture nervose.
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Keefer, Edward W. "Unique applications of cultured neuronal networks in pharmacology, toxicology, and basic neuroscience." Thesis, University of North Texas, 2001. https://digital.library.unt.edu/ark:/67531/metadc2797/.
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This dissertation research explored the capabilities of neuronal networks grown on substrate integrated microelectrode arrays in vitro with emphasis on utilizing such preparations in three specific application domains: pharmacology and drug development, biosensors and neurotoxicology, and the study of burst and synaptic mechanisms. Chapter 1 details the testing of seven novel AChE inhibitors, demonstrating that neuronal networks rapidly detect small molecular differences in closely related compounds, and reveal information about their probable physiological effects that are not attainable through biochemical characterization alone. Chapter 2 shows how neuronal networks may be used to classify and characterize an unknown compound. The compound, trimethylol propane phosphate (TMPP) elicited changes in network activity that resembled those induced by bicuculline, a known epileptogenic. Further work determined that TMPP produces its effects on network activity through a competitive inhibition of the GABAA receptor. This demonstrates that neuronal networks can provide rapid, reliable warning of the presence of toxic substances, and from the manner in which the spontaneous activity changes provide information on the class of compound present and its potential physiological effects. Additional simple pharmacological tests can provide valuable information on primary mechanisms involved in the altered neuronal network responses. Chapter 3 explores the effects produced by a radical simplification of synaptic driving forces. With all synaptic interactions pharmacologically limited to those mediated through the NMDA synapse, spinal cord networks exhibited an extremely regular burst oscillation characterized by a period of 2.9 ± 0.3 s, with mean coefficients of variation of 3.7, 4.7, and 4.9 % for burst rate, burst duration, and inter-burst interval, respectively (16 separate cultures). The reliability of expression of this oscillation suggests that it may represent a fundamental mechanism of importance during periods of NMDA receptor dominated activity, such as embryonic and early postnatal development. NMDA synapse mediated activity produces a precise oscillatory state that allows the study of excitatory-coupled network dynamics, burst mechanisms, emergent network properties, and structure-function relationships.
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De, Faria Da Fonseca Bárbara. "Regulation of Mesenchymal Differentiation Potentials in the avian Neural Crest." Thesis, Paris 6, 2017. http://www.theses.fr/2017PA066146/document.
Full textAbstract:
La crête neurale (CN) est une structure multipotente transitoire de l'embryon de vertébrés. La CN céphalique (CNC), mais pas la CN troncale (CNT), fournit des tissus mésenchymateux (squelette, derme et tissus adipeux de la face). Cette capacité de la CNC est liée à l'absence d'expression des gènes de type Hox. Cependant, les cellules de la CNT possèdent des potentialités mésenchymateuses à l'état dormant, qui peuvent s'exprimer en culture. Les mécanismes moléculaires qui régulent les potentialités mésenchymateuses de la CN le long de l'axe antéro-postérieur restent incompris. Chez l'embryon d'oiseau, nous avons étudié l'influence des facteurs de transcription Hox et Six sur la formation du mésenchyme par la CN. D'une part, nos analyses in vivo et in vitro montrent que Six1 est présent dans des cellules mésenchymateuses de la CN et du mésoderme, suggérant un rôle dans le développement musculo-squelettique de la tête. D'autre part, nous avons testé l'hypothèse d'un rôle inhibiteur des facteurs Hox. Nos résultats montrent que l'expression ectopique de Hoxa2 dans les cellules de CNC en culture inhibe la production d'ostéoblastes, sans affecter celle des cellules nerveuses et mélanocytaires. Dans la CNT, nous avons trouvé que la différentiation osseuse, cartilagineuse et adipocytaire, est fortement réduite après la surexpression de Hoxa2, sans effet sur les autres phénotypes dérivés de la CN. Ces résultats suggèrent que les potentialités mésenchymateuses de la CN sont régulées, au moins en partie, par un mécanisme commun aux cellules de CNC et CNT, mettant en jeu une inhibition de l'activité du gène Hoxa2The neural crest (NC) is a transitory multipotent structure of the vertebrate embryo. The cephalic NC (CNC), not the trunk NC (TNC), gives rise to mesenchymal cell types (contributing to craniofacial skeleton, dermis and adipose tissue). This capacity of the CNC has been linked to the absence of Hox gene expression in the most rostral region of the embryo. However, TNC cells do have mesenchymal potentialities, although in a dormant state in vivo, but which can be disclosed after NC in vitro culture. The molecular mechanisms that regulate mesenchymal potentials of the NC cells along the rostral-caudal axis are still elusive. Here, we have used the avian embryo model to investigate the possible influence on NC mesenchymal fate, of Hox and Six transcription factor genes. On the one hand, in vivo and in vitro culture analyses show that Six1 gene is expressed in mesenchymal cell populations derived from both cranial NC and mesoderm, suggesting a role for Six1 in muscle-skeletal development in the head. On the other hand, we have tested the hypothesis of an inhibitory action of Hox genes on NC cell mesenchymal differentiation using NC in vitro cultures. In CNC cells, we found that ectopic expression of Hoxa2 strongly reduces the production of osteoblasts, while neural and melanocytic phenotypes are unaffected. In the cultured CNT cells, overexpression of Hoxa2 results in largely impaired differentiation into bone cells, chondrocytes and adipocytes, whereas other NC derivatives are unchanged. These results suggest that mesenchymal potentials of the CNC and TNC are controlled, at least in part, via a common mechanism that involves inhibition of Hoxa2 gene activity
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Santa, Maria Cara L. "Optimization of Cell Culture Procedures for Growing Neural Networks on Microelectrode Arrays." Thesis, University of North Texas, 2007. https://digital.library.unt.edu/ark:/67531/metadc5126/.
Full textAbstract:
This thesis describes the development of an optimized method for culturing dissociated, monolayer neuronal networks from murine frontal cortex and midbrain. It is presented as a guidebook for use by cell culture specialists and laboratory personnel who require updated and complete procedures for use with microelectrode array (MEA) recording technology. Specific cell culture protocols, contamination prevention and control, as well common problems encountered within the cell culture facility, are discussed. This volume offers value and utility to the rapidly expanding fields of MEA recording and neuronal cell culture. Due to increasing interest in determining the mechanisms underlying Parkinson's disease, the newly developed procedures for mesencephalon isolation and culture on MEAs are an important research contribution.
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Dian, Emese Emöke. "Application of Cultured Neuronal Networks for Use as Biological Sensors in Water Toxicology and Lipid Signaling." Thesis, University of North Texas, 2004. https://digital.library.unt.edu/ark:/67531/metadc5557/.
Full textAbstract:
This dissertation research explored the capabilities of neuronal networks grown on substrate integrated microelectrode arrays in vitro to be applied to toxicological research and lipid signaling. Chapter 1 details the effects of chlorine on neuronal network spontaneous electrical activity and pharmacological sensitivity. This study demonstrates that neuronal networks can maintain baseline spontaneous activity, and respond normally to pharmacological manipulations in the present of three times the chlorine present in drinking water. The findings suggest that neuronal networks may be used as biological sensors to monitor the quality of water and the presence of novel toxicants that cannot be detected by conventional sensors. Chapter 2 details the neuromodulatory effects of N-acylethanolamides (NAEs) on the spontaneous electrical activity of neuronal networks. NAEs are a group of lipids that can mimic the effects of marijuana and can be derived from a variety of plant sources including soy lecithin. The most prominent NAEs in soy lecithin, palmitoylethanolamide (PEA) and linoleoylethanolamide (LEA), were tested individually and were found to significantly inhibit neuronal spiking and bursting activity. These effects were potentiated by a mixture of NAEs as found in a HPLC enriched fraction from soy lecithin. Cannabinoid receptor-1 (CB1-R) antagonists and other cannabinoid pathway modulators indicated that the CB1-R was not directly involved in the effects of NAEs, but that enzymatic degradation and cellular uptake were more likely targets. The results demonstrate that neuronal networks may also be a viable platform for the elucidation of biochemical pathways and drug mechanisms of action.
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Johnston, D. A. "The avian neural crest : behaviour and long-term survival in culture." Thesis, University of Southampton, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.376464.
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Rogers, A. T. "Spinal cord cell culture : a model for neuronal development and disease." Thesis, University of Bath, 1988. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.234048.
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Johnston, Helen Barbara. "An investigation of aluminium neurotoxicity using in vitro neural culture systems." Thesis, University of Hertfordshire, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.358304.
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Jones, Erin Boote. "Effects of substrate and co-culture on neural progenitor cell differentiation." [Ames, Iowa : Iowa State University], 2008.
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Karbanová, Jana, Tomáš Soukup, Jakub Suchánek, Robert Pytlík, Denis Corbeil, and Jaroslav Mokrý. "Characterization of Dental Pulp Stem Cells from Impacted Third Molars Cultured in Low Serum-Containing Medium." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-136045.
Full textAbstract:
We isolated and expanded stem cells from dental pulp from extracted third molars using an innovative culture method consisting of low serum-containing medium supplemented with epidermal growth factor and platelet-derived growth factor BB. We evaluated the differentiation potential of these cells when they were growing either adherently or as micromass/spheroid cultures in various media. Undifferentiated and differentiated cells were analyzed by flow cytometry, immunocytochemistry and immunoblotting. The flow cytometry results showed that the dental pulp stem cells (DPSCs) were positive for mesenchymal stromal cell markers, but negative for hematopoietic markers. Immunocytochemical and/or immunoblotting analyses revealed the expression of numerous stem cell markers, including nanog, Sox2, nestin, Musashi-1 and nucleostemin, whereas they were negative for markers associated with differentiated neural, vascular and hepatic cells. Surprisingly, the cells were only slightly positive for α-smooth muscle actin, and a heterogeneous expression of CD146 was observed. When cultured in osteogenic media, they expressed osteonectin, osteopontin and procollagen I, and in micromass cultures, they produced collagen I. DPSCs cultured in TGF-β1/3-supplemented media produced extracellular matrix typical of cartilaginous tissue. The addition of vascular endothelial growth factor to serum-free media resulted in the expression of endothelial markers. Interestingly, when cultured in neurogenic media, DPSCs exhibited de novo or upregulated markers of undifferentiated and differentiated neural cells. Collectively, our data show that DPSCs are self-renewing and able to express markers of bone, cartilage, vascular and neural tissues, suggesting their multipotential capacity. Their easy accessibility makes these cells a suitable source of somatic stem cells for tissue engineeringDieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich
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Xydas, Dimitris. "Investigation of spatiotemporal state dynamics in neuronal cultures." Thesis, University of Reading, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.558807.
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Biological neuronal cultures offer the unique opportunity to study the emergent behaviour of networks comprised of a large number of neurones within a controlled environment. Such neurones typically develop random interconnections, yet exhibit certain spontaneous properties reflective of their original function. In vitro cultures are maintained and ob- served using specialised multi-electrode arrays (MEAs), which record their activity in the form of extracellular voltages called spikes and are able to concurrently stimulate them at multiple locations via electrical impulses. Currently, cultures are often being used as the primary controllers in biological-machine hybrids. One of the most intriguing questions however is how to best capture the com- plex interactions of neuronal ensembles and their evolution in time, which would lead to a better understanding of mechanisms of learning and memory. Their spatiotemporal dynamics need to be taken into consideration in any hybrid closed-loop system in order to describe and predict its state at any point in time. To this extent, first the use of temporal coding schemes was examined within spon- taneous and stimulated activity and confirmed certain robust patterns of behaviour. Sub- sequently, both spatial and temporal dynamics of cultures were studied with the use of hidden Markov models (HMMs). Relevant research has demonstrated the capabilities of HMMs in an in vivo context, but their applicability in vitro is not yet widely explored. Two types of models were proposed and important aspects of their framework were anal- ysed for use with multi-channel neuronal recordings. The models were used to analysed a range of spontaneous activity and evoked responses and uncovered robust, repeated dynamic patterns. This led to an important question: how do these dynamic states relate between different conditions? Spectral clustering techniques were implemented to answer this question and study the extent of similarities between different culture behaviours. Future directions of this work anticipate the application of HMMs to improve real- time systems which utilise neuronal cultures as the controlling brains of artificial robotic agents, with further implications for neuroprosthetics and brain-machine interfaces.
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Johnstone, Alex. "Microfluidic systems for neuronal cell culture." Thesis, University of Nottingham, 2016. http://eprints.nottingham.ac.uk/37822/.
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At a high level of abstraction, the brain is a system for analysing sensory information, and responding appropriately. That information is encoded and stored in the millions of neural circuits that comprise the brain. Deciphering this code is essential to understanding how memories are implemented in physiologically normal brain tissue, and to inferring the nature of some neurological disorders affecting memory such as Alzheimer’s disease, in which the neural encoding is aberrant or unsuccessful. One approach to this problem is to reduce the complexity of the brain functionality to three elements: stimuli, response, and reinforcement. The electrical activity of individual neurons can be recorded with electrodes, capturing the pathways of signal propagation in a network of cells. Individual neurons can be also induced to reliably respond to electrical or optical stimuli, so that they initiate, relay, or even block a signal. If the stimuli to a finite network of cells can be made heterogeneous so that only a sub-population of cells is targeted, then the network can be trained to react in a repeatable way to a given stimulus, testing the concept that the higher order functions of the brain can emerge from a simple set of underlying computational rules. Training however requires a mechanism for reinforcing only some of the possible pathways, in synchrony with stimuli and in response to the recorded network activity. In the intact brain, this mechanism is pharmacological: a neuromodulator such as dopamine is released throughout the brain, but as it only coincides with some but not all neuronal activity, the reinforcement is temporally selective. The key task of this project is to emulate this selective neuromodulator reinforcement in vitro in a finite neuronal network. The project must also provide capacity for heterogeneous stimulation and individual cell recording, which can be coordinated with the reinforcement under computer control. The strategy used was to develop microscale chambers to house a small network of cultured neurons. The chambers were integrated with existing cell recording and stimulating technologies, so that specific connections between neurons could be both monitored and induced. Neuronal cultures of a few hundred cells were successfully grown in microchannels, on substrates capable of recording their electrical activity. Thus it was possible to create a small cultured network in which complete network activity could be detected, subject to a sufficiently precise recording technique. Additionally, a fluid-handling system was developed in order to emulate the continual replenishment of nutrients and soluble gases that are essential to cell survival. The system is intended to deliver soluble chemicals that modulate neuronal activity, on a timescale that is consistent with neuromodulator delivery in the body. The fluid handling system comprises a set of pressure driven pumps under automated computer control. This system has the capacity to deliver neuromodulator in solution with high spatiotemporal precision. The ability to reliably deliver and wash off precise volumes of drugs in a matter of seconds, with no dilution of the intended concentration, will be of great benefit to researchers investigating the response of various cell types to different agonists.
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Konagaya, Shuhei. "Design of cell culture substrates for large-scale preparation of neural cells." 京都大学 (Kyoto University), 2013. http://hdl.handle.net/2433/174963.
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Santa, Maria Cara L. Gross Guenter W. "Optimization of cell culture procedures for growing neural networks on microelectrode arrays." [Denton, Tex.] : University of North Texas, 2007. http://digital.library.unt.edu/permalink/meta-dc-5126.
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Jiang, Xiaosong. "Muscle induces neuronal expression of acetylcholinesterase in neuron-muscle co-culture : transcription regulation mediated by cAMP-dependent signaling /." View Abstract or Full-Text, 2003. http://library.ust.hk/cgi/db/thesis.pl?BIOL%202003%20JIANG.
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Thesis (M. Phil.)--Hong Kong University of Science and Technology, 2003.Includes bibliographical references (leaves 132-149). Also available in electronic version. Access restricted to campus users.
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Karbanová, Jana, Tomáš Soukup, Jakub Suchánek, Robert Pytlík, Denis Corbeil, and Jaroslav Mokrý. "Characterization of Dental Pulp Stem Cells from Impacted Third Molars Cultured in Low Serum-Containing Medium." Karger, 2011. https://tud.qucosa.de/id/qucosa%3A27697.
Full textAbstract:
We isolated and expanded stem cells from dental pulp from extracted third molars using an innovative culture method consisting of low serum-containing medium supplemented with epidermal growth factor and platelet-derived growth factor BB. We evaluated the differentiation potential of these cells when they were growing either adherently or as micromass/spheroid cultures in various media. Undifferentiated and differentiated cells were analyzed by flow cytometry, immunocytochemistry and immunoblotting. The flow cytometry results showed that the dental pulp stem cells (DPSCs) were positive for mesenchymal stromal cell markers, but negative for hematopoietic markers. Immunocytochemical and/or immunoblotting analyses revealed the expression of numerous stem cell markers, including nanog, Sox2, nestin, Musashi-1 and nucleostemin, whereas they were negative for markers associated with differentiated neural, vascular and hepatic cells. Surprisingly, the cells were only slightly positive for α-smooth muscle actin, and a heterogeneous expression of CD146 was observed. When cultured in osteogenic media, they expressed osteonectin, osteopontin and procollagen I, and in micromass cultures, they produced collagen I. DPSCs cultured in TGF-β1/3-supplemented media produced extracellular matrix typical of cartilaginous tissue. The addition of vascular endothelial growth factor to serum-free media resulted in the expression of endothelial markers. Interestingly, when cultured in neurogenic media, DPSCs exhibited de novo or upregulated markers of undifferentiated and differentiated neural cells. Collectively, our data show that DPSCs are self-renewing and able to express markers of bone, cartilage, vascular and neural tissues, suggesting their multipotential capacity. Their easy accessibility makes these cells a suitable source of somatic stem cells for tissue engineering.Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.
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Gómez, Orlandi Javier. "Noise, coherent activity and network structure in neuronal cultures." Doctoral thesis, Universitat de Barcelona, 2015. http://hdl.handle.net/10803/346925.
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In this thesis we apply a multidisciplinary approach, based on statistical physics and complex systems, to the study of neuronal dynamics. We focus on understanding, using theoretical and computational tools, how collective neuronal activity emerges in a controlled system, a neuronal culture. We show how the interplay between noise and network structure defines the emergent collective behavior of the system.
We build, using theory and simulation, a framework that takes carefully describes spontaneous activity in neuronal cultures by taking into account the underlying network structure of neuronal cultures and use an accurate, yet simple, model for the individual neuronal dynamics. We show that the collective behavior of young cultures is dominated by the nucleation and propagations of activity fronts (bursts) throughout the system. These bursts nucleate at specific sites of the culture, called nucleation points, which result in a highly heterogeneous probability distribution of nucleation. We are able to explain the nucleation mechanism theoretically as a mechanism of noise propagation and amplification called noise focusing.
We also explore the internal structure of activity avalanches by using well--defined regular networks, in which all the neurons have the same connectivity rules (motifs). Within these networks, we are able to associate to the avalanches an effective velocity and topological size and relate it to specific motifs. We also devise a continuum description of a neuronal culture at the mesoscale, i.e., we move away from the single neuron dynamics into a coarse--grained description that is able to capture most of the characteristic observables presented in previous chapters. This thesis also studies the spontaneous activity of neuronal cultures within the framework of quorum percolation. We study the effect of network structure within quorum percolation and propose a new model, called stochastic quorum percolation, that includes dynamics and the effect of internal noise.
Finally, we use tools from information theory, namely transfer entropy, to show how to reliably infer the connectivity of a neuronal network from its activity, and how to distinguish between different excitatory and inhibitory connections purely from the activity, with no prior knowledge of the different neuronal types. The technique works directly on the fluorescence traces obtained in calcium imaging experiments, without the need to infer the underlying spike trains.
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Albutt, Darren James. "Surface chemistry modification of glass and gold for low density neural cell culture." Thesis, University of Nottingham, 2013. http://eprints.nottingham.ac.uk/13823/.
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Surface chemical modifications are presented for supporting primary neurons in culture. The initial substrates for culture were glass and gold. The surface modifications were based on self assembled monolayer (SAM) approaches. Glass surfaces were initially modified by silanisation with either 3-aminopropyltrimethoxysilane (APTMS) or 3-aminopropyldimethylethoxysilane (APDES), to present amino-terminated surfaces. Gold surfaces were initially modified by thiol SAMs of either 11-amino-1-undecanethiol (AUT) or a peptide fragment of laminin (PA22-2), to present an amino- or peptide-terminated surface respectively. The amine-terminated surfaces of both glass and gold were subject to further modification. A heterobifunctional linker, containing a polyethylene glycol (PEG) spacer, was used to couple the peptide PA22-2 to the amino-terminated surfaces. Surface modifications were characterised using WCA, XPS and ToF-SIMS. The heterobifunctional linker bound homogeneously across the AUT SAM surface, however the linker was not distributed evenly on either of the amino silanisations of glass. Primary neurons from dissociated embryonic rat hippocampi were cultured on the modified glass and gold surfaces. The cell viability was measured during a 3 week long culture using calcein and ethidium homodimer fluorescence. Neuronal cell cultures were viable on all the gold surface modifications. The only viable glass surface was a control surface of adsorbed poly-l-lysine (PLL) on glass. Cell viability on the AUT and the Peptide-PEG-AUT modified gold surfaces was equivalent to the PLL coated glass. Inclusion of the PEG linker reduced protein adsorption from the media to the peptide modified gold surface, allowing cells to recognise the peptide rather than an adsorbed protein layer and improving their viability. The presented gold surface modifications provide suitable substrates for neural cultures which can be used in existing applications for investigating neural activity, such as; multi-electrode arrays, micro-fluidics devices, and surface plasmon resonance.
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Fonseca, Erika Toledo da. "Identificação de áreas neurogênicas no sistema nervoso central de cobaias (Cavia porcellus, Linnaeus 1758): cultura, caracterização e diferenciação de precursores neurais." Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/10/10132/tde-04072013-120221/.
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O presente trabalho relata a identificação de áreas neurogênicas, o cultivo, identificação e caracterização de precursores neurais obtidos do encéfalo de fetos de cobaias. Áreas potencialmente neurogênicas no encéfalo de neonatos foram identificadas por imunohistoquímica para a bromodeoxiuridina (BrdU), após inoculação da droga. A incorporação de BrdU foi detectada principalmente em células das áreas subjacentes ao ventrículo lateral, hipocampo e bulbo olfatório, indicando proliferação celular e sugerindo o potencial neurogênico dessas áreas. A seguir, realizou-se o cultivo de precursores neurais a partir da zona subventricular (SVZ) dos ventrículos laterais. Após aproximadamente uma semana de cultivo, as células da SVZ em multiplicação ativa originaram abundantes massas celulares (neuroesferas, NSFs). Células dissociadas a partir das NSFs primárias foram capazes de gerar novas neuroesferas após alguns dias de cultivo. As NSFs proliferaram em número e em tamanho, podendo ser subcultivadas por até 5-6 passagens, com intervalos de uma semana, permanecendo viáveis por até 60 dias. Nesse período, as NSFs foram congeladas e descongeladas, tendo sido preservadas a sua viabilidade e capacidade proliferativa. Ensaios de viabilidade pelo método colorimétrico de MTT revelaram diferenças de viabilidade entre NSFs cultivadas a partir de SVZs de animais de diferentes idades (fetos, 7, 30 e 180 dias de vida). NSFs dissociadas apresentaram cerca de 2,3% de morte celular por apoptose e cerca de 3,1% de morte por necrose. NSFs íntegras e dissociadas foram submetidas a imunofluorescência (IF), utilizando-se anticorpos para marcadores de células-tronco (nestina), neurônios (Beta-III-tubulina), oligodendrócitos (mGALC) e astrócitos (GFAP). Marcação difusa, de intensidade variável, foi observada no citoplasma de células das NSFs e nas células individualizadas, mas o percentual de células expressando os diferentes marcadores não foi determinado. Quando cultivadas em meio contendo B27, sem fatores de crescimento, as NSFs apresentaram evidências de diferenciação, originando células aderentes a superfície do frasco, com características morfológicas diferentes as das NSFs, indicando diferenciação. Citometria de fluxo de células das NSFs após a diferenciação revelou 13,3% positivas para nestina, aproximadamente 5,5% positivas para beta-III-tubulina, 9% positivas para GFAP e 7,8% positivas para mGalC. Células diferenciadas foram então submetidas a teste de funcionalidade, pela mensuração de influxo de cálcio após estímulo com ácido gama amino butírico (GABA) e glutamato. A adição de GABA e glutamato resultou em estimulação de algumas células diferenciadas, que foi observado por meio do aumento da fluorescência das mesmas. Portanto, células cultivadas e diferenciadas in vitro a partir da SVZ de fetos de cobaias apresentam indicadores funcionais de neurônios. A capacidade das células da SVZ de fetos cobaias originarem células neurais funcionais in vitro é promissora no sentido de aprofundar as pesquisas e viabilizar o uso terapêutico de células-tronco em disordens do sistema nervoso.The present study concerns the identification of neurogenic areas, culture, identification and characterization of neural precursors obtained from the brain of guinea pigs. Potentially neurogenic areas in the brain of neonates were identified by immunohistochemistry for bromodeoxyuridine (BrdU), following administration of the drug. BrdU incorporation was detected mainly in cells underlying the lateral ventricle, in hippocampus and olfactory bulbs, indicating cell proliferation and suggesting a neurogenic potential for these areas. Subsequently, neural precursors were cultured from cells obtained from the subventricular zone (SVZ) of the lateral ventricles. After approximately one week culture, SVZ cells in active multiplication originated abundant cellular masses (neurospheres, NSFs). Cells dissociated from primary NSFs were capable of originating new NSFs after a few days of culture. NSFs proliferated in number and size, allowing 5-6 weekly subculturings and maintaining growth and viability for up to 60 days. In the meantime, NSFs were frozen and thawed, maintaining the viability and proliferative ability. Viability assays by the colorimetric MTT revealed viability differences among NSFs originated from SVZs from animals of different ages (fetuses, 7, 30 and 180 days of age). Dissociated NSFs underwent approximately 2.3% cell death by apoptosis and 3.1% death by necrosis. Intact and dissociated NSFs were submitted to immunofluorescence (IF), using antibodies for cell markers of stem cells (nestin), neurons (beta-III tubulin), oligodendrocytes (mGalC) and astrocytes (GFAP). A diffuse staining of variable intensity was observed in the cytoplasm of NSFS and dissociated cells, yet the rate of cells expressing each individual marker could not be determined. Upon culture in medium containing B27, without NSFs-specific growth factors, NSFs displayed evidence of neural differentiation, originating cells with morphology distinct from that of NSFs, suggesting differentiation. Flow cytometry analysis of NSFs cells after differentiation revealed approximately 13.3% positive for nestin, around 5.5% positive for beta-III-tubulin, 9% GFAP positive and approximately 7.8% positive for mGalC. Differentiated cells were then submitted to a functional test, by measuring calcium influx upon gamma butiric amino acid (GABA) and glutamate stimuli. GABA and glutamate stimulated some differentiated cells. Thus, cells cultured and differentiated from guinea pig SVZ present physiological indicators of neuronal physiology. Thus, the ability of guinea pig SVZ cells to originate functional neurons in vitro is promising towards further studies and potential therapeutic use of neural stem cells in disorders of the nervous system.
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Cēbers, Gvido. "Modulation of AMPA glutamate receptor functions in primary neuronal cultures /." Stockholm, 1999. http://diss.kib.ki.se/1999/91-628-3424-X/.
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Fardet, Tanguy. "Growth and activity of neuronal cultures : emergence of organized behaviors." Thesis, Sorbonne Paris Cité, 2018. http://www.theses.fr/2018USPCC002/document.
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Dans cette thèse, je propose plusieurs modèles et outils numériques afin de mieux comprendre et prédire le comportement et le développement de cultures et dispositifs neuronaux.Les cultures de neurones ont en effet été un outil précieux durant les 20 dernières années : elles ont permis de mieux comprendre la manière dont le cerveau traite les différentes informations qui lui parviennent en donnant aux scientifiques la possibilité de tester les effets de médicaments sur les neurones, ainsi que d'obtenir leurs réponses détaillées à diverses perturbations et stimuli.De plus, de récentes avancées en microfluidiques ont ouvert la voie à la conception de dispositifs neuronaux plus élaborés, rapprochant encore un peu plus la perspective du traitement de signaux complexes via des neurones in vitro.Dans une première partie, je propose un mécanisme pour expliquer les bouffées d'activité épileptiformes présentes dans les cultures, mécanisme que je formule via un modèle théorique concis. J'effectue ensuite une vérification expérimentale des prédictions du modèle sur des cultures et montre que celles-ci sont effectivement compatibles avec le comportement observé in vitro.Dans une seconde partie, je décris plus en détail la description de la dynamique spatio-temporelle du phénomène, notamment le fait que les bursts nucléent en des zones bien précises du réseau neuronal.Comme les prédictions et analyses effectuées dépendent fortement de la structure de ce réseau, je présente ensuite la réalisation d'une plateforme de simulation afin de permettre de modéliser efficacement le développement des réseaux neuronaux. Ce logiciel prend en compte les interactions entre les neurones et leur environnement et constitue la première plateforme à fournir des modèles polyvalents et complets pour décrire l'intégralité du processus de croissance neuronal. Je montre ensuite que ce simulateur est capable de générer des morphologies valides et l'utilise pour proposer des nouvelles topologies de réseaux afin de décrire les cultures de neurones. Je reproduis également des dispositifs neuronaux existants et montre que les activités entretenues par ces structures sont compatibles avec les observations expérimentales. Enfin, je discute plusieurs directions de recherche possibles, pour lesquelles l'utilisation de dispositifs neuronaux spécifiques permettrait de contourner les limitations des cultures neuronales et fournirait ainsi de nouvelles informations sur les processus sous-tendant le développement et la plasticité cérébraleIn this thesis, I provide models and numerical tools to better understand and predict the behavior and development of neuronal cultures and devices.Neuronal cultures have proven invaluable in improving our understanding of how the brain processes information, by enabling researchers to investigate neuronal and network response functions to various perturbations and stimuli.Furthermore, recent progress in microfluidics have opened the gate towards more elaborated neuronal devices, bringing us one step closer to complex signal processing with living in vitro neurons.In a first part, I propose a mechanism to explain the epileptiform bursts of activity present in cultures, mechanism which I formulate as a concise theoretical model. I subsequently test the predictions of this model on cultures and show that they are indeed compatible with the behavior observed in vitro.I further develop this description in the second part of the thesis, where I analyze its spatiotemporal dynamics and the fact that burst nucleate in specific areas in the network.Since predictions and analysis of these nucleation centers strongly depends on the network structure, I develop a simulation platform to enable efficient modeling of the network development. This software takes into account the interactions between the neurons and their environment and is the first platform to provide versatile and complete models to simulate the entire growth process of neurons. I demonstrate that this simulator is able to generate valid neuronal morphologies, then use it to propose new network topologies to describe neuronal cultures, as well as to reproduce existing neuronal devices. I then show that the activities sustained by these structures are compatible with the experimental recordings.Eventually, I discuss several future directions for which the use of neuronal devices would enable to circumvent current limitations of neuronal cultures, thus providing new information on the processes which underlie brain development and plasticity
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Kershaw, Timothy Richard. "The development and transplantation of neural cell lines from the H-2K'b-tsA58 transgenic mouse." Thesis, King's College London (University of London), 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.244043.
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Chao, Zenas C. "Toward the neurocomputer goal-directed learning in embodied cultured networks/." Diss., Atlanta, Ga. : Georgia Institute of Technology, 2007. http://hdl.handle.net/1853/19816.
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Thesis (Ph.D)--Biomedical Engineering, Georgia Institute of Technology, 2008.Committee Chair: Potter, Steve; Committee Member: Butera, Robert; Committee Member: DeMarse, Thomas; Committee Member: Jaeger, Dieter; Committee Member: Lee, Robert.
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Joshi, Pranav. "Three-Dimensional Human Neural Stem Cell Culture for High-Throughput Assessment of Developmental Neurotoxicity." Cleveland State University / OhioLINK, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=csu155965254496159.
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Shi, Xinglong. "DIFFERENTIATION OF NEURAL STEM CELL USING SMALL MOLECULES IN 2D AND 3D CULTURE SYSTEM." Master's thesis, Temple University Libraries, 2015. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/363660.
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BioengineeringM.S.
The neuronal differentiation of neural stem cells (NSCs) has received much attention due to its potential for the treatment of neurodegenerative diseases (i.e., Parkinson’s and Alzheimer’s diseases). In this regard, discovering compounds that direct differentiation of NSCs is highly required to facilitate therapeutic applications. In this study, we examined various bioactive compounds (SA1, SA2, LiCl, compound B, and DHED) to induce the neuronal differentiation of human neural stem cells (hNSCs). The study was conducted on the cells grown in three dimensional (3D) hydrogel or two dimensional (2D) environment since 3D hydrogel mimics the extracellular matrix and provides physiologically more relevant environment than 2D cell culture system. Three-dimensional (3D) hydrogel systems in this study involve polysaccharides such as alginate and hyaluronic acid. Neuronal differentiation of hNSCs was monitored in genetic level and protein level by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunocytochemistry (ICC), respectively. This study will show the effect of bioactive compounds on hNSCs differentiation in 2D and 3D culture systems.
Temple University--Theses