PMID: 7513788May 1, 1994Paper

How variable is a spontaneous mutation rate in cultured mammalian cells?

Mutation Research
J J BoesenJ W Simons

Abstract

The Luria-Delbrück fluctuation analysis provides a method to estimate mutation rates and is commonly applied in somatic cell genetics and in cancer biology. We developed an assay for a Luria-Delbrück fluctuation analysis using the mouse lymphoma cell line, GRSL13. As these cells grow in suspension, one can handle hundreds of parallel cultures using multiwell dishes and dispensers. This assay thereby allows not only an accurate determination of the mutation rate per cell generation but also makes it possible to determine at which time after seeding mutations take place. Using approx. 8000 parallel cultures it has been possible to test whether the mutation rate is constant during the assay. It has been found that the spontaneous mutation rate of GRSL13 cells decreases in the course of a fluctuation test from 2 x 10(-6) to about 2 x 10(-7)/cell/generation. It was shown that this increased replication fidelity may partly be caused by cell density: maintenance of cells at high cell density resulted in a spontaneous mutation rate of 0.7 +/- 4.0 x 10(-7) compared to 4.0 +/- 3.1 x 10(-7) for the standard protocol. In contrast, growing the cells at extremely low cell density resulted in an enhanced mutation rate of 7.7 +/- 1.3 x 10(-7)....Continue Reading

References

Nov 1, 1979·Somatic Cell Genetics·S Goldberg, V Defendi
Oct 1, 1976·Science·P C Nowell
Aug 1, 1992·Molecular & General Genetics : MGG·J J BoesenJ W Simons
May 1, 1990·Mutation Research·L A Loeb, K C Cheng
Jan 1, 1989·Teratogenesis, Carcinogenesis, and Mutagenesis·B Kaina
Dec 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·M R CapecchiG M Wahl
Jan 1, 1973·Mutation Research·R L Capizzi, J W Jameson
Nov 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·M A Cifone, I J Fidler

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Citations

Nov 12, 2009·Regenerative Medicine·Nathalie LefortMarc Peschanski
Sep 19, 2001·Reviews in the Neurosciences·K Heininger
Dec 11, 2014·JMIR Research Protocols·Michael BordonaroTobias May
Apr 30, 1998·Genetics·B S Strauss
Dec 23, 2008·Mutation Research·Xiaowei WuMarek Kimmel

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