Cellular impedance biosensor for drug testing on HCT-116 cancer cells /
Conventional in vitro cytotoxicity methods such as Sulforhodamine B (SRB) and tetrazolium salt (MTT) assays have several limitations such as being expensive, labor-intensive, complicated, toxic, and having discrete measurements and giving single-end results. To this extent, cell-based impedance bios...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
Kuala Lumpur :
Kulliyyah of Engineering, Internationai Islamic University Malaysia,
2015
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| Subjects: | |
| Online Access: | http://studentrepo.iium.edu.my/handle/123456789/4391 |
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| Summary: | Conventional in vitro cytotoxicity methods such as Sulforhodamine B (SRB) and tetrazolium salt (MTT) assays have several limitations such as being expensive, labor-intensive, complicated, toxic, and having discrete measurements and giving single-end results. To this extent, cell-based impedance biosensor has gained huge attention as an alternative to the routinely used methods to study cell growth and spreading by real-time monitoring, and continuous measurement. Impedance biosensor is also inexpensive and uncomplicated. The objectives of this study were to develop an impedance biosensor for detecting the impedance of human colon cancer, HCT-116 cells and determine the changes in cell behavior when treated with anti-cancer agent, 5-Fluorouracil (5-FU), compared to the untreated one; thus investigating the potential of the impedance biosensor in drug testing applications. The electrode fabrication process was done using Paint software and MEMs technology. The impedance of the HCT-116 cells monolayer was detected by Agilent 4294A Precision Impedance Analyzer at frequency ranges from 40 Hz to 10 MHz for every 8 hour intervals in 120 hours. The biosensor consists of two electrodes; working electrode (WE) and counter electrode (CE). To have better cell attachment on the electrode's surface, it was coated with extracellular matrix (ECM); gelatin and polyaniline. From the study, polyaniline gave the best cell attachment at 40 Hz frequency with high impedance measurement, ~ 4250 ohm rather than ~1500 ohm with gelatin. Cytokinetic behavior had indicated that 5-FU inhibited the HCT-116 cells at IC50 of 6.8 µg/ mL. By Trypan blue method, the cell concentrations was reduced approximately 5% when treated with 2.5 µg/ mL, and 10% when treated with 6.8 µg/ mL, both are compared to the untreated one. Results for the constructed impedance biosensor are comparable to the Trypan blue method since both gave similar cell growth trend. It can be concluded that the impedance biosensor has potential to be used as an alternative method in drug testing applications. |
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| Physical Description: | xv, 120 leaves : ill. ; 30cm. |
| Bibliography: | Includes bibliographical references (leave 102-113). |