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Abstract
This study has been carried out to better understand how cells behave in three dimensional polymer scaffolds for cell-based assay applications. Many whole cell-based assays in use today rely on flat, two-dimensional (2D) glass or plastic substrates that may not produce results characteristic of in vivo conditions. Ideal cell-based screening systems call for research efforts to create simple, robust and effective 3D cell-based platforms so that cellular responses will be more representative of those under in vivo conditions. Although 3-D cell culture systems are known to reflect the in vivo behavior of many cell types and are promising approaches for advanced drug screening, providing an appropriate environment in which to culture cells in three dimensions is no easy matter. A major reason that the 3-D culture systems have not entered the drug screening process to date is the lack of simple, controlled techniques and protocols for rapid, standardized 3-D cell-based assay systems. The overall objective of this study is to design a more physiologically relevant cell-based assay system by integrating cells with synthetic polymer scaffolds. A technology was invented to integrate 3D synthetic polymer scaffolds with standard cell culture vessels. This technology can be used to feasibly modify any traditional 2D cell-based assay vessels for 3D cell-based assay with currently used high throughput screening (HTS) systems. Extensive research has been conducted to compare cellular activities on polymer scaffolds (3-D), flat surfaces (2-D) and in vivo surrogates. To best mimic the natural extracellular matrix, nano-structures have been brought into the previous scaffold topography to form a nano-fibrous and micro-porous combination scaffolds. Results obtained from this study supports conclusion that cells cultured on 3D scaffolds more closely emulates in-vivo surrogates such as fresh dissected tissue and neural spheres, in comparison to the cells on flat (2-D) controls. This new biomimetic cell-based assay platform may provide a broadly applicable 3D cell-based system for use in drug discovery programs and other research fields. Future studies could demonstrate the potential of this 3-D cell-based assay system in improving clinical efficacy and drug safety prediction in early stages of drug discovery programs.