Skip to main content
Top
Published in: Radiation Oncology 1/2012

Open Access 01-12-2012 | Methodology

Determination of cell survival after irradiation via clonogenic assay versus multiple MTT Assay - A comparative study

Authors: Karl Buch, Tanja Peters, Thomas Nawroth, Markus Sänger, Heinz Schmidberger, Peter Langguth

Published in: Radiation Oncology | Issue 1/2012

Login to get access

Abstract

For studying proliferation and determination of survival of cancer cells after irradiation, the multiple MTT assay, based on the reduction of a yellow water soluble tetrazolium salt to a purple water insoluble formazan dye by living cells was modified from a single-point towards a proliferation assay. This assay can be performed with a large number of samples in short time using multi-well-plates, assays can be performed semi-automatically with a microplate reader. Survival, the calculated parameter in this assay, is determined mathematically. Exponential growth in both control and irradiated groups was proven as the underlying basis of the applicability of the multiple MTT assay. The equivalence to a clonogenic survival assay with its disadvantages such as time consumption was proven in two setups including plating of cells before and after irradiation. Three cell lines (A 549, LN 229 and F 98) were included in the experiment to study its principal and general applicability.
Appendix
Available only for authorised users
Literature
1.
go back to reference Pauwels B, Korst AE, de Pooter CM, Pattyn GG, Lambrechts HA, Baay MF, Lardon F, Vermorken JB: Comparison of the sulforhodamine B assay and the clonogenic assay for in vitro chemoradiation studies. Cancer Chemother Pharmacol 2003, 51: 221-226.PubMed Pauwels B, Korst AE, de Pooter CM, Pattyn GG, Lambrechts HA, Baay MF, Lardon F, Vermorken JB: Comparison of the sulforhodamine B assay and the clonogenic assay for in vitro chemoradiation studies. Cancer Chemother Pharmacol 2003, 51: 221-226.PubMed
2.
go back to reference Saunders MI: Predictive testing of radiosensitivity in non-small cell carcinoma of the lung. Lung Cancer 1994,10(Suppl 1):S83-90.CrossRefPubMed Saunders MI: Predictive testing of radiosensitivity in non-small cell carcinoma of the lung. Lung Cancer 1994,10(Suppl 1):S83-90.CrossRefPubMed
3.
go back to reference Williams JR, Zhang Y, Zhou H, Gridley DS, Koch CJ, Slater JM, Little JB: Overview of radiosensitivity of human tumor cells to low-dose-rate irradiation. Int J Radiat Oncol Biol Phys 2008, 72: 909-917. 10.1016/j.ijrobp.2008.06.1928CrossRefPubMed Williams JR, Zhang Y, Zhou H, Gridley DS, Koch CJ, Slater JM, Little JB: Overview of radiosensitivity of human tumor cells to low-dose-rate irradiation. Int J Radiat Oncol Biol Phys 2008, 72: 909-917. 10.1016/j.ijrobp.2008.06.1928CrossRefPubMed
4.
go back to reference Eble MJ, Hensley FW, Flentje M, Schlotz A, Wannenmacher M: A modified computer-assisted colorimetric microtitre assay (MTT) to assess in vitro radiosensitivity of V79, CaSki, HeLa and WiDr cells. Int J Radiat Biol 1994, 65: 193-201. 10.1080/09553009414550231CrossRefPubMed Eble MJ, Hensley FW, Flentje M, Schlotz A, Wannenmacher M: A modified computer-assisted colorimetric microtitre assay (MTT) to assess in vitro radiosensitivity of V79, CaSki, HeLa and WiDr cells. Int J Radiat Biol 1994, 65: 193-201. 10.1080/09553009414550231CrossRefPubMed
5.
go back to reference Kratzke RA, Kramer BS: Evaluation of in vitro chemosensitivity using human lung cancer cell lines. J Cell Biochem Suppl 1996, 24: 160-164.CrossRefPubMed Kratzke RA, Kramer BS: Evaluation of in vitro chemosensitivity using human lung cancer cell lines. J Cell Biochem Suppl 1996, 24: 160-164.CrossRefPubMed
6.
go back to reference Foldbjerg R, Dang DA, Autrup H: Cytotoxicity and genotoxicity of silver nanoparticles in the human lung cancer cell line, A549. Arch Toxicol 2010. Foldbjerg R, Dang DA, Autrup H: Cytotoxicity and genotoxicity of silver nanoparticles in the human lung cancer cell line, A549. Arch Toxicol 2010.
7.
go back to reference Hansen J, Bross P: A cellular viability assay to monitor drug toxicity. Methods Mol Biol 2010, 648: 303-311. 10.1007/978-1-60761-756-3_21CrossRefPubMed Hansen J, Bross P: A cellular viability assay to monitor drug toxicity. Methods Mol Biol 2010, 648: 303-311. 10.1007/978-1-60761-756-3_21CrossRefPubMed
8.
go back to reference Price P, McMillan TJ: Use of the tetrazolium assay in measuring the response of human tumor cells to ionizing radiation. Cancer Res 1990, 50: 1392-1396.PubMed Price P, McMillan TJ: Use of the tetrazolium assay in measuring the response of human tumor cells to ionizing radiation. Cancer Res 1990, 50: 1392-1396.PubMed
9.
go back to reference Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C: Clonogenic assay of cells in vitro. Nat Protoc 2006, 1: 2315-2319. 10.1038/nprot.2006.339CrossRefPubMed Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C: Clonogenic assay of cells in vitro. Nat Protoc 2006, 1: 2315-2319. 10.1038/nprot.2006.339CrossRefPubMed
10.
go back to reference Price P, McMillan TJ: The use of non-clonogenic assays in measuring the response of cells in vitro to ionising radiation. Eur J Cancer 1994, 30A: 838-841.CrossRefPubMed Price P, McMillan TJ: The use of non-clonogenic assays in measuring the response of cells in vitro to ionising radiation. Eur J Cancer 1994, 30A: 838-841.CrossRefPubMed
Metadata
Title
Determination of cell survival after irradiation via clonogenic assay versus multiple MTT Assay - A comparative study
Authors
Karl Buch
Tanja Peters
Thomas Nawroth
Markus Sänger
Heinz Schmidberger
Peter Langguth
Publication date
01-12-2012
Publisher
BioMed Central
Published in
Radiation Oncology / Issue 1/2012
Electronic ISSN: 1748-717X
DOI
https://doi.org/10.1186/1748-717X-7-1

Other articles of this Issue 1/2012

Radiation Oncology 1/2012 Go to the issue