PMID: 2111457May 1, 1990Paper

A quantitative assay for measuring the induction of mutations in human peripheral blood T-lymphocytes

Mutation Research
T NorimuraJ J McCormick

Abstract

We optimized conditions for propagating freshly-isolated human peripheral blood T-lymphocytes and cells that had been stored in liquid nitrogen on Day 5 post-isolation, exposing them to mutagens in exponential growth, and measuring the cytotoxicity of the agent from the loss of colony-forming ability, and its mutagenicity from the increase in frequency of 6-thioguanine-resistant cells. Supernatant containing T-cell growth factor, from 60Co-irradiated peripheral mononuclear cells cultured in the presence of 60Co-irradiated B-lymphoblastoid human cells as allogeneic stimulators, supplied at a concentration of 10% along with 10% serum and 10(5) allogeneic stimulator cells/ml, supported exponential growth (population doubling times of 22 h) for extended periods (greater than 30 d). It gave cloning efficiencies of greater than or equal to 40%. T-lymphocytes stored in liquid nitrogen and returned to culture shortly before mutagen exposure exhibited the same sensitivity as freshly-isolated T-cells to killing by the agents tested, i.e., UV radiation, ethyl-nitrosourea, and (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,19 alpha,epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene. We showed that if mutagenized populations frozen during the expression peri...Continue Reading

References

Aug 1, 1986·Mutation Research·A W SkulimowskiM Haliandros
Aug 1, 1984·Mutation Research·B J SandersonA A Morley
Mar 10, 1983·Nature·A A MorleyR G Ryall
Nov 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·R J AlbertiniW R Borcherding

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