Spelling suggestions: "subject:"well microenvironment"" "subject:"well microenvironments""
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Interakce buňek s biomateriály v tkáňovém inženýrství tvrdých a měkkých tkání / Cell-biomaterial interactions in hard and soft tissue engineeringZárubová, Jana January 2016 (has links)
Tissue engineering is an interdisciplinary field which aims to create substitutes of damaged tissues by combining cells with biomaterials. Cells are extremely sensitive to their microenvironment and so the cell response to biomaterials can be regulated by different extrinsic stimuli and alterations of biomaterial properties. Successful implant integration into the tissue can therefore be promoted by appropriate surface roughness, chemical composition, adhesion ligand density, as well as the availability of growth factors. This thesis mainly focuses on the development of orthopedic replacements and the improvement of the currently used blood vessel prostheses. Through the study of cell-biomaterial interactions, it was demonstrated that superimposed topography with features ranging from the nano to micro scale promotes cell spreading, proliferation, and the metabolic activity of osteoblast-like cells. Moreover, when comparing the chemical composition of biomaterials for orthopedic implants, higher osteoblast densities were observed on composites with 5-15 vol. % of calcium phosphate nanoparticles, while concentrations of 25 vol. % did not support cell proliferation. Cell viability, however, was not affected. In vivo, a more intensive formation of new bone tissue, was found on samples containing...
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Microengineered Substrates for Systematic Probing Of Cardiomyocytes’ Morphology, Structure, and FunctionJamilpour, Nima, Jamilpour, Nima January 2017 (has links)
The inability of the myocardium to regenerate after injury plus the inadequate number of available hearts for transplantation have drawn attention to the creation of functional tissue constructs for implantation within the injured heart. In addition, there is an increasing interest in developing in vitro models to study heart physiology and pathology as well as to evaluate drug efficacy. Formation of these in vitro models and tissue constructs requires highly specific conditions to mimic the normal environment of cells in the body. Firstly, in this study, plasma lithography patterning of elastomeric substrates is exploited for creating microtissues composed of neonatal cardiomyocytes, and investigating their development in different mechanical microenvironments. Immunofluorescence microscopy and force spectroscopy show that the size and shape of the cardiomyocyte clusters, as well as the sarcomere length, fiber alignment, and beating amplitude and frequency of the cardiomyocytes, are regulated by microenvironmental cues. Computational analysis reveals that the mechanical stress at the cluster-substrate interface strongly correlates with the aforementioned characteristics of the cardiomyocytes. Taken together, our results underscore a collective mechanoadaptation scheme in cardiac development. Secondly, a silicone substrate with tunable elasticity is characterized for biological studies. Uniaxial tensile testing and microindentation show that these substrates could cover the biological range of stiffness for normal and pathological conditions. Spectrophotometry demonstrates that the transmittance of these substrates is comparable to those of glass and Sylgard 184. Atomic force microscopy shows that the surface roughness of samples is lower than that of widely-used Sylgard 184. Contact angle measurements before and after exposure to air plasma indicate that these samples are compatible with plasma lithography patterning. Thirdly, a new technique for cell patterning is developed which utilizes selective plasma lithography to modify protein adhesion on the substrate. This approach is based on controlling the conformation of Pluronic F-127 layer adsorbed on the surface by modifying surface wettability. Contact angle measurements show that both PDMS and plastic petri dish are compatible with this technique. X-ray photoelectron spectroscopy and atomic force microscopy confirm the adsorption of PF-127 layers with controlled conformation. Fluorescent and bright-field microscopy demonstrate selective adhesion of proteins and attachment of cells merely on plasma-treated areas. Finally, micropillar arrays are employed to determine the effects of two proteins associated with regulation of thin filament length, i.e. Lmod2 and Tmod1, on contractile force generation at the cellular level. Our results demonstrate that the contractile force of single isolated Lmod2-KO cardiomyocytes decreases compared to the wildtype control. Transduction of Lmod2 in the knockout cardiomyocytes restores their contractile force to the level of their WT counterparts, verifying that the observed contractile dysfunction is specific to the loss of Lmod2. Our data demonstrate that overexpression of Tmod1 in cardiomyocytes decreases their contractile force compared to the WT cells and confirm the effects of Lmod2 knockout on contractile force generation.
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Orthogonal Click Chemistry Hydrogels for Culture and Differentiation of Pluripotent Stem CellsMatthew R Arkenberg (13021746) 08 July 2022 (has links)
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<p>Pluripotent stem cells (PSCs) are increasingly utilized to investigate early human developmental processes including gastrulation and organogenesis of endoderm-derived pancreatic lineages. Critical for tissue development, the PSC niche is a dynamic environment consisting of extracellular matrix (ECM) components that guide cell proliferation, migration, and differentiation. However, investigation of the interplay between the PSC niche and organogenesis has been limited to conventional two-dimensional (2D) cell culture or three-dimensional (3D) platforms requiring use of ill-defined materials (e.g., Matrigel). Furthermore, these systems lack tunability to probe specific qualities of the PSC niche including mechanical properties and biochemical compositions. In this dissertation, modular and dynamic hydrogels were designed to study PSC and niche interactions during differentiation and pancreatic organogenesis. Specifically, two bioorthogonal chemical reactions, thiol-norbornene photopolymerization and tetrazine-norbornene inverse electron demand Diels-Alder (iEDDA) reactions were employed to generate gelatin- and poly(ethylene glycol) (PEG)-based hydrogels with spatiotemporally tunable physicochemical properties. Following mechanical characterization of the hydrogels, the multicomponent gelatin-based hydrogels were assessed for supporting viability and pluripotency of human induced pluripotent stem cells (hiPSCs), as well as for permitting their trilineage differentiation. Next, fully synthetic PEG-based hydrogels with temporally tunable crosslinking density were established to probe the effect of matrix mechanics on definitive endoderm differentiation of the hiPSCs. Finally, hiPSC-to-pancreatic progenitor cell differentiation was explored in both naturally-derived gelatin-based hydrogels and synthetic PEG-based hydrogels, with cells differentiated on a 2D surface as a control. Overall, this work demonstrates that culture dimensionality, material compositions, and mechanics profoundly influence hiPSC differentiation and pancreatic morphogenesis.</p>
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MICRO/NANOSCALE ENGINEERING OF THE CELL MICROENVIRONMENTGallego-Perez, Daniel 28 July 2011 (has links)
No description available.
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New microfluidic systems for controlling the cell microenvironment during live-cell imaging / Développement de systèmes microfluidiques pour des applications biologiques sous microscopie haute résolutionBabic, Julien 14 December 2017 (has links)
Connaître en temps réel la réponse et le comportement des cellules et organismes modèles suite à des changements de leur environnement, ou à des modulations de leurs fonctions biologiques est devenu essentiel dans les sciences du vivant. Ces réponses nous permettent ensuite de comprendre les mécanismes qui régissent le fonctionnement des cellules vivantes, avec des implications en recherche fondamentale, appliquée et biomédicale. Un des plus gros défis technologiques reste le contrôle des paramètres environnementaux en microscopie haute résolution. De nos jours, aucun système ne permet de réguler un ensemble complexe de paramètres de manière précise, dynamique et simultanée tout en observant les cellules dans leur environnement. L’objectif de ma thèse est de mettre au point un tel dispositif permettant a minima une régulation fine de la température, de la composition du milieu, et notamment de la concentration de divers drogues. Ce système doit être compatible avec les applications les plus poussées en microscopie photonique. Mon approche au cours de ma thèse pour élaborer un tel système est l’utilisation de la microfluidique. En effet, c’est la seule technologie qui puisse de réaliser un tel multiplexage. Elle permet de manipuler des petites quantités de fluide à travers un système contenant des canaux de dimensions allant du micromètre au centimètre. Cet ordre de grandeur des canaux constitue un atout majeur (réduction de la consommation des réactifs, réduction des couts, cinétiques des réactions chimiques et biologiques élevées, temps de diffusion court, etc.) et permet d’allier les expériences biologiques à la microscopie. Mon objectif est de concevoir une puce microfluidique qui représentera un pas technologique majeur et ouvrira de nouvelles possibilités de recherche. / Monitoring in real-time the response of cells and model organisms to the changes in their environment or to modulations of their biological functions has become essential in life sciences. One of the main technical challenges for biologists is the precise and dynamic control of various environmental parameters while doing high-resolution microscopy. My thesis consists of building a robust and versatile system, dedicated to live-cell imaging that will be compatible with adherent and non adherent models, that could provide a precise and simultaneous control of 1) the temperature, 2) the media exchanges and 3) the drug concentration while doing photonic microscopy. My approach is to use microfluidics, which is the best candidate in order to achieve this system and provides all the necessary controls of micro-scaled volumes for culturing, maintaining or analyzing cells. It produces miniaturized systems used as tools for biological experiments, in which channels of a micro-scaled dimension are used for the fluid circulation. The laminar flow in these chips allows fast molecule diffusion as well as fast temperature diffusion. Because of the high surface to volume ratio, the consumption of reagents is reduced, and media switches can be fast. This system will represent a major technical and beneficial step and will open new possibilities of research in biology.
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Adult stem cells in the trachea and tracheal submucosal glandsLynch, Thomas John 01 August 2016 (has links)
Breathing is essential for human life, yet tens of millions of people in the U.S. alone suffer from lung diseases. With each breath, lungs are exposed to the external environment. Inhaled air first passes through the trachea, bronchi, and finally the bronchioles before it reaches the alveoli where gases are exchanged. A barrier of epithelial cells protects the airways. In addition, epithelial glands also secrete protein-rich fluids onto the airway surfaces to help maintain sterility. Injury, disease, or other factors can damage these cells, and regiospecific stem cells (SCs) can divide to replace them. However, many important details about lung SCs are still unknown. For example, what processes control SC division? How do region-specific SCs differ from one another? And how does disease or injury impact SC biology? We found that some processes that regulate lung development also control adult SC division following injury. We show that SCs from airway glands give rise to surface epithelial cell types and glandular cell types. In contrast, surface SCs only generated surface cell types. Finally, we identify a type of cell in the glands that can regenerate surface cell types after severe injury. These studies provide new insights into the neighborhoods in which SCs reside in the large airways and processes that control their contribution to airway repair following injury. Overall, this research provides important new insights into adult SC biology and conditions affecting lung health.
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Micro-engineering of embryonic stem cells niche to regulate neural cell differentiationJoshi, Ramila, Joshi January 2018 (has links)
No description available.
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