L’une des principales sources d’inconfort dans un hélicoptère sont les vibrations transmises par le rotor à la structure de l’appareil. En vol d’avancement, des efforts aérodynamiques cycliques sont subis par l’ensemble des pales en tête rotor et génèrent de très fortes vibrations basse fréquence (aux alentours des 17Hz) transmises aux passagers via la boîte de transmission principale puis le fuselage lui-même. Afin de garantir le confort des membres d’équipage et des passagers, de nombreux systèmes antivibratoires ont été conçus. Ces systèmes sont généralement passifs car la majorité de l’énergie vibratoire transmise à la structure se situe à une fréquence unique ωc correspondant à bΩ avec b le nombre de pales et Ω la fréquence de rotation du rotor. Cependant, les appareils modernes évoluent et le régime rotor jusqu’alors fixe durant toutes les phases de vol varie à présent pour des préoccupations de performances et de consommation (variation de l’ordre de +/-10% autour de bΩ). Cette nouvelle contrainte dans la conception des hélicoptères rend pertinente la technologie des systèmes antivibratoires actifs, pouvant s’adapter à la sollicitation en termes d’amplitude et fréquence. Lors de ces travaux de thèse, la suspension passive SARIB de Airbus Helicopters basée sur le principe du DAVI (Dynamic Antiresonant Vibration Isolator) est modifiée afin d’être rendue active par ajout d’une partie actuation/commande. La théorie des lois et algorithmes de contrôle utilisés dans ces travaux, est présentée en détail afin de poser solidement les bases du contrôle actif du prototype de suspension conceptualisé ici à savoir le contrôle FXLMS (adaptatif) et le contrôle optimal LQG. Afin de simuler le fonctionnement du système, un modèle tridimensionnel de la suspension active est construit, couplé à la structure souple de l’hélicoptère (NH90). Sur ce modèle sont alors appliquées les différentes lois de commande introduites auparavant et leurs performances comparées dans différents cas de chargement en tête rotor et surtout pour différentes fréquences de sollicitation. De même, pour chaque algorithme, différentes localisations des capteurs d’erreur sont étudiées afin de converger vers une configuration optimale. Les simulations démontrent que l’algorithme FXLMS feedforward est très bien adapté au contrôle des perturbations harmoniques et permet de réduire très significativement le niveau vibratoire du plancher cabine, sans réinjection parasite dans le reste de la structure. Une comparaison de l’efficacité du SARIB actif avec les systèmes d’absorbeurs en cabine est ensuite effectuée pour démontrer la pertinence d’utiliser le principe du DAVI comme base d’un système actif. Les travaux de cette thèse traitent également des essais réalisés en laboratoire sur le prototype échelle 1 de la suspension SARIB active avec contrôle FXLMS. / One of the main causes of discomfort in helicopters are the vibrations transmitted from the rotor to the structure. In forward flight, the blades are submitted to cyclic aerodynamic loads which generate low frequency (around 17Hz) but high energy mechanical vibrations. These vibrations are transmitted from the rotor to the main gearbox, then to the structure and finally to the crew and passengers. In order to maintain acceptable comfort for crew members and passengers, a lot of antivibration devices have been developed since the last 30 years. These systems are generally passive because most of the mechanical energy transmitted to the structure is at only one frequency ωc which is equal to the product bΩ with b the number of blades and Ω the rotor rotational speed. However, modern helicopters evolve and the rotor rpm, which has always been considered as fixed during flight is now a function of time, depending on the flight phases in order to increase performances and reduce energy consumption (variation bandwidth of Ω +/- 10%). This new constraint on the design of helicopters makes the active antivibration technology completely relevant with its capacity to adapt in terms of amplitude and frequency to the perturbation. During this thesis, the passive suspension called SARIB from Airbus Helicopters, based on the DAVI principle (Dynamic Antiresonant Vibration Isolator) is modified in order to implement active components and command (actuation). The theory of the control algorithms used in this thesis is presented in detail in order to define the theoretical tools of the active DAVI control which are : FXLMS control (adaptive control) and LQG (optimal control). To simulate the complete system, a 3D multibody model of the active suspension has been set up, coupled to a the flexible structure of a NH90 (Airbus Helicopters). On this model are applied the different control algorithms presented before and their performances are compared for different loads with variable frequency on the rotor hub. In the same way, different locations for the error sensors in the structure are studied to find the optimal control configuration. The simulations show that the FXLMS algorithm is well suited for the control of harmonic perturbations and reduce significantly the dynamic acceleration level on the cabin floor, without parasite reinjection on other parts of the structure. A comparison of the active SARIB with classical cabin vibration absorbers is also made in terms of efficiency in order to show the advantages of using the DAVI system as a base for an active antivibration device. Finally, this thesis also presents the experiments realized in the dynamics laboratory of Airbus Helicopters on a 1:1 scale prototype of the active SARIB suspension with FXLMS control. The results demonstrate the efficiency of the active suspension architecture and control algorithms.

L’objectif de cette étude est de bien décrire les mécanismes impliqués dans la remise en suspension des microparticules en conduite ventilée. Une méthodologie expérimentale est utilisée, et cela en tenant compte de la période d’accélération de l’écoulement qui précède l’atteinte du régime permanent. Une méthode optique a été choisi pour étudier le mouvement initial des particules, et leur cinétique de remise en suspension. Parallèlement, nous avons recueilli des données locales de l’écoulement en période d’accélération et au régime permanent.Le démarrage de la remise en suspension a été analysé relativement à l’évolution temporelle de trois paramètres pouvant influencer le démarrage de la remise en suspension, le premier est la vitesse instantanée en proche paroi, le second est l’intensité turbulente, et enfin le troisième est l’énergie cinétique turbulente. Le paramètre prépondérant sur le démarrage de la remise en suspension semble être l’énergie cinétique turbulente. / The objective of this study is to well describe the mechanisms involved in the resuspension of particles in ventilated duct by using an experimental methodology and taking into account the acceleration of the air flow which always precedes steady state. For that purpose, we chose an optical method in order to investigate the initial movement of particles, and to quantify the resuspension kinetics. In parallel we collected local data of the flow during acceleration and steady state.We observed that the resuspension kinetics starts during the acceleration period and extends to steady state. We highlighted the relevant velocity characteristics (critical velocity at the center duct and close to the wall, critical kinetic energy range) to explain this phenomenon. The resuspension start seems to be linked with a critical kinetic energy range.

Memoria (licenciado en ciencias jurídicas y sociales) / En el primer capítulo de este estudio comenzaremos estudiando brevemente y en forma general el contrato de trabajo y los derechos y obligaciones que derivan de éste. Sólo algunos de estos derechos y obligaciones pueden suspenderse sin dar término a la relación laboral y serán el legislador, las partes o el tribunal, en su caso, quienes determinen de que forma puede llevarse a cabo dicha suspensión y los efectos que se producirán. Una vez determinado el punto mencionado anteriormente nos abocaremos al estudio del concepto y naturaleza jurídica de la suspensión de la relación laboral. En el segundo capítulo abordaremos el ámbito internacional. En primer lugar examinaremos los aspectos relativos a la Organización Internacional del Trabajo (O.I.T.) de la cual han emanado diversos instrumentos internacionales sobre los más diversos temas laborales y de seguridad social que han inspirado el avance legislativo de la mayoría de los países del mundo. Si bien no existen documentos específicos relativos a la suspensión, de este organismo han emanado convenios y recomendaciones sobre algunas materias que constituyen causales de suspensión de la relación laboral, las cuales examinaremos brevemente para obtener las directrices de este organismo sobre tales materias. En segundo lugar analizaremos el derecho comparado en materia de suspensión. Estudiando el derecho laboral de la mayoría de los países de Hispanoamérica por separado determinaremos la tendencia en esta materia: si la suspensión se regula en cada país analizado; en caso de estar reglamentada, cuáles son las causales que se consideran y los efectos que se contemplan; y en caso de no estar contemplada en forma sistemática, cuáles son causas que a nuestro juicio constituyen suspensión. En este punto puede parecer que la investigación es demasiado extensa, pero lo consideramos necesario porque es la mejor forma de obtener una visión clara y comparativa entre las distintas legislaciones. Desde esta perspectiva, y dada la amplitud de materias que se tratan, creemos que esta memoria es un aporte en el estudio de los más distintos temas laborales en derecho comparado, y que marca la diferencia con publicaciones anteriores sobre el tema, al analizar legislaciones extranjeras en forma detallada y al contemplar países que no suelen considerarse en estas investigaciones. En el capítulo tercero estudiaremos exhaustivamente la suspensión de la relación laboral en la legislación chilena. Como esta institución no se encuentra regulada en forma sistemática en la legislación nacional, analizaremos los casos que a nuestro juicio constituyen causales de suspensión. Otras publicaciones anteriores sobre este tema acotan el concepto de suspensión y restringen el estudio de sus causales. Por nuestra parte, hemos optado por abordar este tema de la manera más amplia posible, de forma tal que nuestro análisis sea completo y desprejuiciado, incluyendo las últimas modificaciones a la legislación laboral. Para un estudio sistematizado de cada uno de los casos considerados dividiremos el estudio de éstos en suspensión legal, convencional y judicial. En el último capítulo de este estudio analizaremos la jurisprudencia relativa a la suspensión de la relación laboral, tanto la administrativa como la judicial.

University of Macau / Faculty of Science and Technology / Department of Electromechanical Engineering

Kyoto University (京都大学) / 0048 / 新制・課程博士 / 博士(工学) / 甲第14589号 / 工博第3057号 / 新制||工||1455(附属図書館) / 26941 / UT51-2009-D301 / 京都大学大学院工学研究科化学工学専攻 / (主査)教授 山本 量一, 教授 宮原 稔, 教授 大嶋 正裕 / 学位規則第4条第1項該当

Abstract Over recent years the progress in actuator and microelectronics technology has made intelligent suspension systems feasible. These systems are designed to reduce the drivers' exposure to harmful vibration, as well as to improve the handling properties of the vehicle. Due to widespread use of vehicles as an example of a true MIMO-system, a myriad of different control schemes and algorithms can be found in the literature for these systems. Linearized models are commonly used when the control algorithms are derived. This thesis describes the development of a new analytical full vehicle model, which takes the essential kinematics of the suspension system into account, as well as a new approach to controlling the full vehicle vibration problem. The method of calculating the desired damping forces for each of the semi-active actuators is based on the skyhook theory and this new model is introduced. The performance of the control schemes is evaluated with simulations in a virtual environment. For the excitation to the vehicle, standardized ISO-tracks, washboard tracks and single bump tracks were used. The performance between the two different semi-active control systems and the passive system are compared in terms of damping the vibration, variation of the dynamic tire load and demand for rattlespace. The damping of vibration evaluates both the ability to suppress the vibration on heave, pitch and roll degrees of freedom and ability to reduce the drivers' exposure to harmful whole body vibration. The frequency distribution of the vibration was also reviewed. Variation of dynamic tire contact force is evaluated as an RMS-value and the demand for rattlespace is evaluated as a percentage value of the used rattlespace compared to the maximum free stroke provided by the suspension hardware. As a result from this work, the theory and simulation results are presented. Also a new vehicle model, which takes the essential non-linearity caused by suspension kinematics into account, is presented including all the mathematics needed. The comparison between the passive and the semi-active concepts has been performed on the basis of simulation results. These results show that the novel semi-active concept reduces the driver's exposure to vibration induced by terrain undulations better than any earlier proposed version. Also variation of dynamic tire load is reduced with a novel concept, while it suffers a drawback in the demand for the rattlespace.

Magister Pharmaceuticae - MPharm / Co-crystals are a solid phase phenomena that could enhance the physicochemical properties of an active pharmaceutical ingredient. A co-crystal has never been incorporated into a liquid dosage form with the assurance of maintaining its co-crystal state until absorption under defined conditions. This study aims to develop a liquid formulation with a nevirapine co-crystal. A protocol was developed to investigate all the five co-formers that were used to make the nevirapine co-crystals to-date. The most appropriate co-former was selected for a liquid dosage form to study the integrity and the scaling up of the co-crystal in a suspension formulation. Co-formers used were viz. saccharin, glutaric acid, salicylic acid, rac-tartaric acid and maleic acid. These were characterized according to their physical, chemical, pharmacological and pharmaceutical properties. A grading scale was used to select the most appropriate co-former for a suspension formulation. Comparatively, saccharin produced the best combination of physical, chemical, pharmacological and pharmaceutical properties, especially with regard to the particle size and the specific gravity which proved to be very useful as optimal criteria for suspension formulation. Upon selection of the ideal co-former, scale-up of the nevirapine saccharin co-crystal was performed from a small scale of 350 mg to a large scale of 5 g. Nevirapine-saccharin (NVSC) co-crystals were prepared utilizing the slow evaporation technique, using methanol as the solvent and the percentage yield of the co-crystals were > 80 %. The identity of co-crystals was confirmed using hot stage microscopy (HSM), differential scanning calorimetry (DSC), fourier transform infra- red (FTIR) and thermogravimetric analysis (TGA). Three co-crystal suspension formulations were prepared using the excipients identified in the branded, Viramune® suspension, with each formulation containing viscosity enhancers such as aerosil 200, carbopol 971G and carbopol 974P. To ascertain the co- crystal integrity in the suspension, it was filtered and the filtrate was identified with DSC and FTIR while the filtered solution was identified with ultraviolet spectroscopy (UV). The co-crystal suspension formulation with optimal pH, viscosity and assurance of co-crystal integrity was the carbopol 974P formulation. The UV and DSC of the filtrate of the suspension revealed that the co-crystal had not separated into its individual components and remained intact while in suspension form irrespective of the excipients added. This formulation proceeded to the quality control stage. It was assessed for its pH, viscosity and dissolution according to the USP 32 standards and compared to the branded nevirapine suspension, Viramune ®, presently on the market. The suspension was characterized for particle size, zeta potential and polydispersity index. The dissolution results assayed by High Performance Liquid Chromatography (HPLC) revealed a drug release of 86 % in the Viramune® suspension while the NVSC co- crystal suspension achieved a drug release of 94% within 30 minutes of dissolution. / National Research Foundation (NRF)

Vibrations caused by rough road excitations influence tracked vehicle dynamic performance. Good capabilities of such vehicles like high mobility, manoeuvrability and comfort are guaranteed by optimal suspension systems. The suspension systems of tracked vehicles are exposed to extreme operating conditions. This creates a conflict between ride comfort and handling that is even greater than the conflict between ride comfort and handling for general road vehicles. Tracked vehicles must be able to traverse not only rough roads but also smooth terrains. The challenges in developing an optimized suspension system for tracked vehicles include the high and changeable damping forces required for tracked vehicles crossing rough terrains. The use of active or semi-active suspension systems overcomes the limitations inherent in the conventional passive suspension. However, active suspension systems are expensive, complicated to design and have high power demand. Thus, semi-active suspension systems have emerged as a good compromise between active and passive suspension system. There is considerable current research on the applications of magnetorheological (MR) fluid dampers for semi-active suspensions of executive brand of some cars. However, there is very little research on semi-active devices for tracked vehicle suspension. In fact, currently, there is no commercially available large scale MR dampers in the market that produce the high damping force to suit such applications. In response to these requirements, this research proposes a novel semi-active tracked vehicle suspension system that uses MR dampers to improve the ride comfort and handling characteristics of tracked vehicles. It also assesses the dynamics of the new suspension with various semi-active control methods. This study is conducted in four phases. The first phase provides a numerical investigation on the dynamic performance of a seven-degrees-of-freedom (7-DOF) passive suspension model of the armour personnel carrier (APC) M113 tracked vehicle. The numerical investigation considers the influence of variation of five suspension design parameters on the vehicle dynamic performance. These parameters include number, locations of hydraulic shock absorber, damping coefficient, suspension and wheel stiffnesses. The results indicate that the optimal suspension performance is attained by using two or three dampers. The best locations for these dampers are at the extreme road wheels i.e. the first, second and last road wheel stations. Moreover, the vehicle performance is reduced when the damping coefficient is increased. Additionally, low suspension stiffness offers better vehicle ride while high wheel stiffness degrades the vehicle performance. These results identify the limitations inherent in the conventional passive suspension. For the second phase, the dynamic characteristics of the hydraulic, hydro-gas and MR dampers are experimentally measured and fitted using the Chebyshev orthogonal functions to produce the restoring force surfaces for each damper, which are compared. On one hand, the restoring force surfaces of the hydraulic and hydro-gas dampers show fixed properties at specified frequencies. On the other hand, the restoring force surfaces of the MR dampers show properties that can be controlled at the same specified frequencies by the variation of the applied current levels. Thus, the potential and the effectiveness of the controllable properties of MR dampers for semi-active vibration control is demonstrated. Also, in this phase, the best set of parameters to use in the modified Bouc-Wen model to characterise the MR dampers, has been derived. The third phase of the project is also experimentally based. A new and novel test rig which represents the 7-DOF scaled suspension model of the tracked vehicle is designed and fabricated. The primary purpose of the test rig is to evaluate the performance of the proposed suspension with MR dampers. Furthermore, experiments are conducted on the test rig to evaluate some semi-active control methods and their effectiveness in reducing suspension vibration. The results show that the use of two or three MR dampers at the extreme wheels offers optimal suspension performance. This confirms the numerical results that are derived from the full scale passive suspension system with hydraulic dampers. The experimental results also show that skyhook control and hybrid control (which combines groundhook and skyhook controls) of the semi-active suspension are more effective in reducing the road-induced vibration and improving the suspension dynamic behaviours. Also, validations of the predicted responses of the semi-active scaled MR suspension model with the measured responses have been presented. The fourth and final phase provides a numerical simulation on the development and evaluation of the semi-active control methods for a full scale tracked vehicle suspension with MR dampers using the validated suspension model. Three semi-active control strategies are proposed. The first two controllers are the skyhook and hybrid controls which provide better suspension performance. In addition, the third controller, which is an intelligent fuzzy-hybrid control system, is used to optimize the suspension performance. The results from this intelligent system are compared with the two traditional control methods (skyhook and hybrid controls) under bump, sinusoidal and random excitations. It is shown that the proposed controller can enhance simultaneously the vehicle ride and handling characteristics.

In this study the feasibility of two mathematical models of the dynamics of a hydraulic bypass valve used on semi-active suspension systems for heavy vehicles is investigated. It is envisaged that similar models will eventually be incorporated into a full vehicle, three dimensional simulation study. The valve system contains an electro-hydraulic pilot valve circuit, a logic element, a damper and four check valves in a rectifier configuration. Models were compiled from first principles in the MATLAB environment and with the commercial fluid power simulation software, AMESim. The numerical methods used in the MATLAB model were found to be incapable of solving the stiff, nonlinear and discontinuous governing equations efficiently, while AMESim is very capable of handling detailed and complex fluid power models. Experimental work was conducted to determine certain steady state model parameters and to obtain dynamic performance data with which to validate model integrity. Several external factors influenced the valve behaviour during experiments making data extraction challenging. Simple first order assumptions accounting for the external influences on the valve therefore had to be included in the models. If this is done the basic dynamic behaviour of the valve system is matched well by the models. In general, the number of unknown parameters associated with fluid power systems accounted for the largest portion of the error between the simulated and measured response. The model as developed proved the possibility of creating highly accurate models but also indicated the amount of effort needed for their compilation. / Dissertation (MEng (Mechanical Engineering))--University of Pretoria, 2007. / Mechanical and Aeronautical Engineering / unrestricted

Induced pluripotent stem cells (iPSCs) have been recognised as a virtually unlimited source of stem cells that can be generated in a patient-specific manner. Due to these cells’ potential to give rise to all differentiated cell types of the human body, they have been widely used to derive differentiated cells for drug screening and disease modelling purposes. iPSCs also garner much interest as they can potentially serve as a source for cell replacement therapy. Towards the realisation of these biomedical applications, this thesis aims to address challenges that are associated with scale-up, safety and biofabrication. Firstly, the manufacture of a high number of human iPSCs (hiPSCs) will require standardised procedures for scale-up and the development of a flexible bioprocessing method, since standard adherent hiPSC culture exhibits limited scalability and is labour-intensive. While the quantity of cells that are required for cell therapy depends largely on the tissue and defect that these replacing cells are meant to correct, an estimate of 1 × 10^9 has been suggested to be sufficient for several indications, including myocardial infarction and islet replacement for diabetes. Here, the development of an integrated, microcarrier-free workflow to transition standard adherent hiPSC culture (6-well plates) to scalable stirred suspension culture in bioreactors (1 L working volume, 2.4 L maximum working volume) is presented. The two-phase bioprocess lasts 14 days and generates hiPSC aggregates measuring 198 ± 58 μm in diameter on the harvesting day, yielding close to 2 × 10^9 cells. hiPSCs can be maintained in stirred suspension for at least 7 weeks with weekly passaging, while exhibiting pluripotency-associated markers TRA-1-60, TRA-1-81, SSEA-4, OCT4, and SOX2. These cells retain their ability to differentiate into cells of all the three germ layers in vitro, exemplified by cells positive for AFP, SMA, or TUBB3. Additionally, they maintain a stable karyotype and continue to respond to specification cues, demonstrated by directed differentiation into beating cardiomyocyte-like cells. Therefore, the aim of manufacturing high hiPSC quantities was met using a state-of-the-art scalable suspension bioreactor platform. Secondly, multipotent stem cells such as induced neural stem cells (iNSCs) may represent a safer source of renewable cells compared to pluripotent stem cells. However, pre-conditioning of stem cells prior to transplantation is a delicate issue to ensure not only proper function in the host but also safety. Here, iNSCs which are normally maintained in the presence of factors such as hLIF, CHIR99021, and SB431542 were cultured in basal medium for distinct periods of time. This wash-out procedure results in lower proliferation while maintaining key neural stem cell marker PAX6, suggesting a transient pre-differentiated state. Such pre-treatment may aid transplantation studies to suppress tumourigenesis through transplanted cells, an approach that is being evaluated using a mouse model of experimental focal demyelination and autoimmune encephalomyelitis. Thirdly, biomedical applications of stem cells can benefit from recent advancements in biofabrication, where cells can be arranged in customisable topographical layouts. Employing a 3DDiscovery bioprinter, a bioink consisting of hiPSCs in gelatin-alginate was extruded into disc-shaped moulds or printed in a cross-hatch infill pattern and cross-linked with calcium ions. In both discs and printed patterns, hiPSCs recovered from these bioprints showed viability of around 70% even after 4 days of culture when loaded into gelatin-alginate solution in aggregate form. They maintained pluripotency-associated markers TRA-1-60 and SSEA-4 and continued to proliferate after re-plating. As further proof-of-principle, printed hiPSC 3D constructs were subjected to targeted neuronal differentiation, developing typical neurite outgrowth and resulting in a widespread network of cells throughout and within the topology of the printed matrix. Staining against TUBB3 confirmed neuronal identity of the differentiated cellular progeny. In conclusion, these data demonstrate that hiPSCs not only survive the 3D-printing process but were able to differentiate along the printed topology in cellular networks. / Induzierte pluripotente Stammzellen (iPSZ) stellen eine praktisch unbegrenzte Stammzellquelle dar, welche patientenspezifisch erzeugt werden kann. Da diese Zellen das Potenzial haben, alle differenzierten Zelltypen des menschlichen Körpers hervorzubringen, werden sie für die Herstellung differenzierter Zellen für Arzneimitteltests und für die Krankheitsmodellierung verwendet. Sie erfahren auch großes Interesse, weil sie als Zellquelle in der Zellersatztherapie Anwendung finden könnten. Die vorliegende Dissertation beschäftigt sich mit drei zentralen Herausforderungen, die im Rahmen der biomedizinischen Anwendung von iPSZ auftreten. Die Herstellung einer großen Zahl von humanen iPSZ (hiPSZ) erfordert die Entwicklung standardisierter Verfahren für die Skalierung, welche durch die Entwicklung einer flexiblen Bioprozessmethode realisiert werden kann. Bisher wird die Skalierbarkeit durch eine standardmäßig adhärente Zellkultur und den damit verbundenen hohen Arbeitsaufwand begrenzt. Die Menge an Zellen, die für die Zelltherapie benötigt wird, hängt stark vom Gewebetyp ab, welcher von den ersetzenden Zellen korrigiert werden soll. Berechnungen legen nahe, dass eine Anzahl 1 × 10^9 Zellen für eine Vielzahl von Indikationen ausreicht – einschließlich Myokardinfarkt und Inselzelltransplantation für Diabetes. Im Rahmen dieser Arbeit wurde ein integrierter Arbeitsablauf zur skalierbaren Zellsuspensionskultur von hiPSZ ohne Verwendung von microcarrier entwickelt, um die standardmäßig adhärente Kultur (6-Well-Platten) in Bioreaktoren (1 L Arbeitsvolumen, 2,4 L maximales Arbeitsvolumen) zu überführen. Der zweiphasige Produktionsprozess dauert 14 Tage und erzeugt hiPSZ-Aggregate mit einem finalen Durchmesser von 198 ± 58 μm, der annähernd 2 × 10^9 Zellen beinhaltet. hiPSZ können mindestens 7 Wochen lang in einer gerührten Zellsuspension bei wöchentlichem Passagieren gehalten werden, wobei sie Pluripotenz-assoziierte Marker wie TRA-1-60, TRA-1-81, SSEA-4, OCT4 und SOX2 beibehalten. Die Zellen behalten weiterhin ihre Fähigkeit, sich in vitro in Zellen mit AFP-, SMA- oder TUBB3-Immunoreaktivität und damit in Zellen aller drei Keimblätter zu differenzieren. Darüber hinaus halten sie einen stabilen Karyotyp aufrecht und reagieren auf gezielt eingesetzte externe Differenzierungsstimuli, wie durch eine gezielte Differenzierung in schlagende Kardiomyozyten-ähnliche Zellen demonstriert werden konnte. Somit wurde das Ziel, eine großen Anzahl hiPSCs herzustellen, mit einer hochmodernen, skalierbaren Suspensionsbioreaktorplattform erreicht. Multipotente Stammzellen wie induzierte neurale Stammzellen (iNSZ) gelten verglichen mit iPSZ als sicherere Zellquelle für Ersatztherapien. Die Vorkonditionierung von Stammzellen vor der Transplantation ist jedoch ein heikles Thema, da sowohl die einwandfreie Funktion im Wirtsgewebe als auch Sicherheit gewährleistet werden müssen. Im Rahmen dieser Arbeit wurden iNSZ, die normalerweise im Kulturmedium mit Faktoren wie hLIF, CHIR99021 und SB431542 gehalten werden, für eine definierte Zeitspanne in basalem Medium kultiviert. Die Vorbehandlung führt zu einer geringeren Proliferation, jedoch unter Erhalt der Expression des wichtigen neuralen Stammzellmarkers PAX6, was auf einen transienten vordifferenzierten Zustand hindeutet. Eine solche Vorbehandlung könnte bei zukünftigen Transplantationsstudien angewandt werden, um die Tumorentstehung durch transplantierte Zellen zu unterdrücken. Dieser Ansatz wird in Zukunft mit einem Mausmodell der experimentellen fokalen Demyelinisierung und der autoimmunen Enzephalomyelitis untersucht. Schließlich kann die Zellersatztherapie von den jüngsten Fortschritten in der Biofabrikation profitieren, bei der die Zellen durch das Drucken in anpassbare topographische Profile angeordnet werden können. Mit einem 3DDiscovery Biodrucker wurde eine Biotinte bestehend aus Gelatine-Alginat und hiPSZ in scheibenförmig extrudiert oder in einem Kreuzschraffurmuster gedruckt und mittels Kalziumionen-Zugabe vernetzt. Gedruckte hiPSZ zeigten auch nach 4 Tagen Kultivierung eine Lebensfähigkeit von etwa 70 % und weiterhin das Auftreten der Pluripotenz-assoziierten Marker TRA-1-60 und SSEA-4. Zudem konnten sie sich anschließend mit standardmäßig adhärenter Zellkultur weiter vermehren. Zudem konnte gezeigt werden, dass die gedruckten Konstrukte einer gezielten neuronalen Differenzierung unterzogen werden können, die zu einem typischen Neuritenauswuchs und zu einer weitreichenden interzellulären Vernetzung durch und innerhalb der Topologie der gedruckten Matrix führte. Die Färbung gegen TUBB3 bestätigte die neuronale Identität der differenzierten Zellen. Zusammenfassend zeigen diese Daten, dass bei Verwendung des in dieser Studie erarbeiteten Protokolls hiPSZ nicht nur den 3D-Druckprozess überleben, sondern auch entlang der gedruckten 3D Topologie in Netzwerke Neurone differenzieren können.