Doctor of Philosophy, The Ohio State University, 2012, Chemical and Biomolecular Engineering

Drug screening is a long and costly process confronted with low productivity and challenges in using animals. 3-D cell-based high-throughput platforms have been developed to improve drug-screening efficiency and minimize animal testing. However, online monitoring of cell proliferation, pH, and dissolved oxygen (DO) has been a challenge in 3-D cell-based assays. In this work, a 40 micro-well plate bioreactor (40-MBR) system was developed as a high-throughput platform for 3-D cell cultures. This 40-MBR has similar dimensions of a 384-well plate (384-MWP) can provide real-time and noninvasive monitoring of cell proliferation, pH, and DO in 3-D microenvironments. A colon cancer cell line expressing enhanced green fluorescent protein (EGFP) under the control of a constitutive CMV promoter was tested with two potential cancer drugs using the 40-MBR and 384-MWP. Compared to the 384-MWP, the 40-MBR gave more reliable and highly reproducible growth kinetic data with reduced experimental errors. This study demonstrated the potential application of the 40-MBR as a high-throughput platform for screening potential cancer drugs and evaluating their cytotoxic effects in the early-stage drug discovery. Cytotoxicity and embryotoxicity of chemicals were also investigated in the 40-MBR using EGFP-expressing embryonic stem cells (ESCs). Embryonic stem cell test (EST) has been used as an in vitro model for assessing embryotoxicity. However, the current EST can only provide end-point data and cannot predict embryotoxicity of chemicals affecting organs such as bone. In this study, a novel high-throughput embryotoxicity assay was developed using EGFP-expressing ESCs under the control of a survivin promoter. Survivin expression is closely associated with embryo development and cell differentiation. For control, ESCs expressing EGFP under the control of a CMV promoter were used to monitor cytotoxicity of chemicals. Using survivin as a diagnostic marker for predicting embryotoxicity was fi (open full item for complete abstract)

Committee: Shang-Tian Yang (Advisor); Jeffery Chalmers (Committee Member); Andre Palmer (Committee Member) Subjects: Chemical Engineering