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VALIDATION OF LUMINESCENT-BASED ENDPOINT ASSAY FOR EVALUATION OF CHEMORESISTANCE IN CANCER
2016 (English)Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
Abstract [en]

Resistance to chemotherapy is one of the major factors responsible for complications and relapses after cancer treatments. Cell viability assays are in vitro assays utilized to foresee efficiency of a specific treatment and thereby avoiding treatments with ineffective chemotherapeutic agents. The purpose of this study was to validate a luminescent-based cell viability assay, CellTiter-Glo® 2.0 assay, and determine whether the method could be incorporated in routine diagnostics at the Department of Clinical Chemistry and Pharmacology (Akademiska laboratoriet, Uppsala, Sweden) replacing the current method, fluorometric microculture cytotoxicity assay (FMCA).To achieve a thorough validation two cell lines; HCT 116 wt and THP-1, with different characteristics were used. In addition the chemotherapeutic agents selected vary in their targeting pathways and are used in treatments for different types of cancer.Prior to its comparison to FMCA the luminescent method underwent optimization with regard to cell seeding density as well as plate type. The methods were compared with respect to the reported cell survival percentages or survival index (SI %).The results varied between the two cell lines with THP-1 SI % values being more or less identical between the two methods where as the luminescence results from HCT 116 wt were systematically lower than those from FMCA.Although the methods seem to yield similar results in certain situations it was determined that it, at this stage, is not enough to conclude that they are interchangeable. Further validation is therefore required with additional substances as well as cell lines to determine the correlation between the two methods.

Place, publisher, year, edition, pages
2016.
Keywords [en]
Chemoresistance, ATP-assay, Survival Index (SI %), Cytotoxicity, Luciferase
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:uu:diva-349036OAI: oai:DiVA.org:uu-349036DiVA, id: diva2:1199280
Educational program
Biomedical Laboratory Science Programme; Biomedical Laboratory Science Programme
Examiners
Available from: 2018-04-20 Created: 2018-04-20 Last updated: 2018-04-20Bibliographically approved

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