BIOMATERIALS FOR THE IMAGE-BASED ANALYSES OF HIGH-THROUGHPUT 3D HEPATIC CULTURES

Abstract

3D spheroid cultures are important to improve the physiological relevance of image data in high content screening. However, existing strategies insufficiently constrain spheroids leading to displacements under indifferent robotic handling, and difficulty in image focusing. Additionally, existing strategies cause poor transparency which interfere with imaging resolution. In this thesis, spheroid-constraining biomaterials in high-throughput configurations are re-designed to confer transparency for improved subcellular imaging resolution, and to facilitate easy image-focusing for improved signal detection and consistency. By controlling polymeric particle size during phase transition of a thermoresponsive polymer, Mie scattering is attenuated and leads to scaffold transparency. This is key to improving subcellular resolution in scaffold-constrained spheroids. By controlling surface physicochemical properties, spheroids can be surface-tethered to offer a level imaging plane for enhanced subcellular signal detection. For long-term studies, in situ transparent macroporous scaffolds offer long-term spheroid stability. For short-term dynamic mechanobiological studies, the tethered spheroid platform offers tunable morphological control. In all, new biomaterial strategies for 3D image-based analyses in high-throughput culture configurations have been achieved.