Abstract:
:A modified protocol is described for the in vitro analysis of micronuclei in whole blood or separated lymphocyte cultures. The induction of binucleate cells by various concentrations of cytochalasin B (3, 4.5 or 6 micrograms/ml) was examined at two harvest times (68 or 72 h). An optimal yield was obtained by adding cytochalasin B at a dose of 6 micrograms/ml to cultures 44 h after initiation with harvest 24 h (whole blood) or 28 h (separated lymphocytes) later. Cytocentrifuge preparations of lymphocytes (separated from whole blood using commercial preparations of Ficoll either at the commencement of the assay or upon harvest) were stained with Acridine Orange. Using this method, cytokinesis-blocked lymphocytes remain intact and micronuclei are readily identified. The method is suitable for both whole blood and separated lymphocyte cultures, thus allowing direct comparisons of sensitivity to genotoxic agents.
journal_name
Mutagenesisjournal_title
Mutagenesisauthors
Ellard S,Parry EMdoi
10.1093/mutage/8.4.317subject
Has Abstractpub_date
1993-07-01 00:00:00pages
317-20issue
4eissn
0267-8357issn
1464-3804journal_volume
8pub_type
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