Generating a fluorescent caspase sensor-based 3D drug screening system

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 la...

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Main Author: Koh, Lin Fang.
Other Authors: School of Chemical and Biomedical Engineering
Format: Final Year Project
Language:English
Published: 2012
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Online Access:http://hdl.handle.net/10356/49914
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-499142023-03-03T15:39:58Z Generating a fluorescent caspase sensor-based 3D drug screening system Koh, Lin Fang. School of Chemical and Biomedical Engineering Luo Qian Kathy DRNTU::Science::Medicine::Biosensors 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. Bachelor of Engineering 2012-05-25T06:34:38Z 2012-05-25T06:34:38Z 2012 2012 Final Year Project (FYP) http://hdl.handle.net/10356/49914 en Nanyang Technological University 59 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Medicine::Biosensors
spellingShingle DRNTU::Science::Medicine::Biosensors
Koh, Lin Fang.
Generating a fluorescent caspase sensor-based 3D drug screening system
description 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.
author2 School of Chemical and Biomedical Engineering
author_facet School of Chemical and Biomedical Engineering
Koh, Lin Fang.
format Final Year Project
author Koh, Lin Fang.
author_sort Koh, Lin Fang.
title Generating a fluorescent caspase sensor-based 3D drug screening system
title_short Generating a fluorescent caspase sensor-based 3D drug screening system
title_full Generating a fluorescent caspase sensor-based 3D drug screening system
title_fullStr Generating a fluorescent caspase sensor-based 3D drug screening system
title_full_unstemmed Generating a fluorescent caspase sensor-based 3D drug screening system
title_sort generating a fluorescent caspase sensor-based 3d drug screening system
publishDate 2012
url http://hdl.handle.net/10356/49914
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