The Use of In Vitro Cultured Lymphocytes for Tracing Mutagenic Activities of Chemicals

Abstract

Lymphocytes from normal, non-smoking human individuals not taking drugs were isolated from the peripheral blood by means of the lymphoprep method. The cells were cultured in RPMI medium with 10% fetal calf serum and stimulated with Phytohemagglutinin. A mutagen such as 3-methylcholanthrene was added for varying periods of time. Then the subspecies of DNA, i.e. double and single stranded DNA (ds-DNA and ss-DNA), were separated by the alkaline elution technique and quantitated by fluorimetric estimation. The mutagen induced a significant rise in the level of ss-DNA, but no changes in ds-DNA could be traced. The time-dependent changes increased for at least four days of exposure, indicating that the repair enzymes were not able to compensate for the DNA damage.

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