engleski

Fast Micromethod as a tool for DNA damage determination in different cell lines and tissues

The induction of DNA damage was measured in native DNA isolated from cotton-spinner Holothuria tubulosa, marine sponge Suberites domuncula cells, mussel mytilus galloprovincialis solid tissue (gills) homogenate, Chinese Hamster Ovary cells (CHO), mouse muscle homogenate, human lymphocytes and human cell line (HeLa cells) by fast Micromethod. This method detects DNA damage (strand breaks, alkali.labile sites and incomplete excission repair) in cell suspension or tissue homogentaes in single microplates. Fast Micromethod requires very short time multiple analysis (less than 3 hours) and less than 10 ng DNA persingle well (about 3000 human, hamster or mouse cells, 12500 Suberites domuncula cells or about 25 ug of mussel gills homogenate). DNA damage could be detected after different doses of gama rays, generated by Cs137 (1800 Ci; 0-1500 rad) and UV-B light 254 nm (0-1000 J/m2) irradiation or chemical treatment by fifferent xenobiotics: NQO (4-nitroquinoline-N-oxide; 0-200 ng/ml), BaP (benzo(a)pyrene; 0-20 ug/g mussel) and Bleomycine (0-83 ug/ml). The method is sensitive to wide range of different doses and concentrations of DNA damaging agents. This makes it applicable for measurement of DNA integrity of small samoles for genotoxicty assessment as well as for analysisi related to human health and occupational exposure.

Fast Micromethod; DNA damage; cell lines; tissue

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