A modified computer-assisted colorimetric microtitre assay (MTT) to assess in vitro radiosensitivity of V79, CaSki, HeLa and WiDr cells

• Verfasst von: Eble, Michael J. [VerfasserIn]
• Verfasst von: Hensley, Frank W. [VerfasserIn]
• Verfasst von: Flentje, Michael [VerfasserIn]
• Verfasst von: Schlotz, Annette [VerfasserIn]
• Verfasst von: Wannenmacher, Michael [VerfasserIn]
• Titel: A modified computer-assisted colorimetric microtitre assay (MTT) to assess in vitro radiosensitivity of V79, CaSki, HeLa and WiDr cells
• Verf.angabe: M.J. Eble, F.W. Hensley, M. Flentje, A. Schlotz, M. Wannenmacher
• Jahr: 1994
• Umfang: 9 S.
• Fussnoten: Gesehen am 06.02.2025 ; Elektronische Reproduktion der Druck-Ausgabe 3. Juli 2009
• Titel Quelle: Enthalten in: International journal of radiation biology
• Ort Quelle: London : Taylor & Francis, 1959
• Jahr Quelle: 1994
• Band/Heft Quelle: 65(1994), 2, Seite 193-201
• ISSN Quelle: 1362-3095
• DOI: doi:10.1080/09553009414550231
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1916564690

Abstract

The non-clonogenic MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, was modified and a new procedure of analysis is proposed. The colorimetric assay could be performed semiautomatically using microtitre plate reader connected to a personal computer. Data are processed and plotted with a customized program. To ensure that both control and irradiated microtitre plates contain exponentially growing cells at the time of analysis, the calculation of relative survival is based on a series of well-defined cell numbers initially seeded. To make the two endpoints of non-clonogenic and clonogenic assays comparable, i.e. counting of living cells versus counting of colonies, radiation-induced progression delay was incorporated into the calculation. Radiation-induced cell killing and progression delay could be determined in a single analysis, but in an independent way. X-ray survival curves were generated for V79, CaSki, WiDr and HeLa cells using the non-clonogenic and a standard clonogenic assay. Using the linear-quadratic formula, the resulting parameters α,β and the mean inactivation dose were not significantly different. The described assay is a feasible and reproducible technique for determination of cellular survival, which may be able to incorporate progression delay. The equivalence to a clonogenic survival assay could be proven.