Generating a fluorescent caspase sensor-based 3D drug screening system
Koh, Lin Fang.
Date of Issue2012
School of Chemical and Biomedical Engineering
A three-dimensional fluorescent biosensor based scaffold was developed using microencapsulation and the feasibility of using this system as a high throughput screening system for anti-cancer drugs was explored. The scaffold was formed by alginate beads containing gelatin, which were coated with a layer of poly-L-lysine. A breast-cancer cell line generated using the fluorescence resonance energy transfer (FRET) had been developed previously which can detect apoptosis by caspase-3 activation in living cells. Observations shown that these MCF-7-C3 biosensor cells embedded within the scaffold remained viable and could proliferate within the bead, forming spheroids of varying sizes. Cell morphology, proliferation and viability were characterized using fluorescent microscopy, MTT assays and the measurement of YFP (Yellow Fluorescent Protein) emissions. Preliminary tests of the effects of an anti-cancer drug, doxorubicin, which is a DNA-intercalating agent, on the cells were also carried out. Experiments to determine the IC50 of the drug on MCF-7-C3 cells plated on flat petri dishes was first conducted before the drug was applied to the spheroids in the bead. However, at this IC50, results showed no significant change in morphology, signifying the higher resistance of the cell spheroids to the drug, which is expected according to previous patterns of anti-cancer drug testing. Thus, the model has the potential to serve as a platform for the screening of chemotherapeutic drugs with further developments such as the application of FRET.
Final Year Project (FYP)
Nanyang Technological University