479-Genetic Toxicology In vitro Sister Chromatid Exchange Assay in Mammalian Cells.pdfVIP

479 Adopted: OECD GUIDELINE FOR TESTING OF CHEMICALS 23 Oct 1986 Genetic Toxicology: In vitro Sister Chromatid Exchange Assay in Mammalian Cells 1. I N T R O D U C T O R Y I N F O R M A T I O N • P r e r e q u i s i t e s – Solid, liquid, vapour or gaseous test substance – Chemical identification of test substance – Purity (impurities) of test substance – Solubility characteristics – Melting point/boiling point – pH (where appropriate) – Vapour pressure data (if available) • S t a n d a r d d o c u m e n t s There are no relevant international standards. 2. M E T H O D A. INTRODUCTION The sister chromatid exchange (SCE) assay is a short-term test for the detection of reciprocal exchanges of DNA between two sister chromatids of a duplicating chromosome. SCEs represent the interchange of DNAreplication products at apparently homologous loci. The exchange process presumably involves DNA breakage and reunion, although little is known about its molecular basis. Detection of SCEs requires some means of differentially labelling sister chromatids, which can be achieved e.g. by incorporation of bromodeoxyuridine (BrdU) into chromosomal DNA for two cell cycles. SCEs can also be measured in mammals and in non-mammalian systems. • P r i n c i p l e o f t h e m e t h o d Mammalian cells in vitro are exposed to thetest chemical with and without an exogenous mammalian metabolic activation system and cultured for two rounds of replication in BrdU- containing medium.After treatment with a spindle inhibitor (e.g. colchicine) to accumulate cells in a metaphas