Feb 1, 1980

A method utilizing human lymphocytes with in vitro metabolic activation for assessing chemical mutagenicity by sister-chromatid exchange analysis

Mutation Research
A D White, L C Hesketh

Abstract

A technique using human lymphocytes together with an Ames-type microsomal (S9) activation system for testing indirect chemical mutagens has been developed and examined. The cytotoxic drug cyclophosphamide (CP), which only displays mutagenic properties after metabolic activation, was used as a test chemical and mutagenicity assessed in terms of sister-chromatid exchange (SCE) induction. Direct exposure of lymphocytes to CP and S9 mix produced increases in the yield of SCE for CP concentrations down to 10(-6) M. Exposure times of 30, 60 and 120 min commencing at the beginning of cell culture or after 48 h gave different ranges of detection and response with later treatment being more effective for SCE induction. Variations in relative proportions of the S9 mix and culture medium constituents affected the lymphocytes' tolerance of toxic factors and modified the mutagen's effect. CP pre-incubated with S9 mix for 1 h before adding to the lymphocyte cultures also produced increases in SCE levels but the method was complicated and did not reduce toxicity. Direct addition of CP and S9 mix to the lymphocytes without prior pre-incubation showed maximum sensitivity, minimum toxicity and was a simpler, more reliable technique.

Mentioned in this Paper

Cyclophosphamide
Lymphocytes as Percentage of Blood Leukocytes (Lab Test)
Cytological Techniques
Sister Chromatid Exchange
Lymphoid Cells
Cell Culture Techniques
Chromatids
Crossing Over, Genetic
Activation, Metabolic
Metabolic Biotransformation

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