Kinetic Analysis of Micro-Physiological Systems Using Automated Live-Cell Imaging
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Chang, Jonathan J.
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Abstract
Micro-physiological systems can be broadly defined as a class of advanced in vitro models which seek to better recapitulate tissue structure and function, and they include technologies such as organoids, organs-on-chips, and 3D tissue constructs. While micro-physiological systems have shown great promise in accelerating preclinical drug development and enhancing mechanistic studies of diseases, challenges in scalability and throughput have limited their utility. Here, three micro-physiological systems were developed which follow a general framework to address these challenges; the micro-physiological systems were formed in commercial 96/384-well plates and combined with automated live-cell imaging to streamline operation and allow for kinetic analysis of cellular behaviors. First, a new method for forming mammary organoids in 384-well ultra-low attachment plates was developed. This system was used to rapidly screen reagent concentrations and optimize for organoid size and circularity. Second, aqueous two-phase systems were utilized to bioprint unprecedentedly small fibrin gels laden with fibroblasts. Fibrinolysis of these gels was determined to be a sensitive and robust correlate of wound healing, and fibrinolysis rate was explored as a functional readout of cellular senescence. Lastly, the fibrin bioprinting technology was adapted to deposit micro-clots over venous endothelial cells. This system was utilized as a disease model to investigate the impact of various drugs and stimuli on the pathogenesis of venous thromboembolism. Overall, the work accomplished in this thesis could help enhance preclinical drug studies and guide future development of micro-physiological systems.
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2023-04-27
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Text
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Dissertation