An efficient and simple method for enriching metaphase cells for dicentric chromosome assay.

Abstract

As compared to peripheral blood mononuclear cell (PBMC) culture, a lower mitotic index (MI) is seen in whole blood (WB) culture, but WB can be directly used for culture in dicentric chromosome assay (DCA). The purpose of this study is to develop a simple protocol for metaphase enrichment to improve the metaphase frequency of WB culture. Fixed cells were obtained after performing WB and PBMC cultures for DCA after conventional fixation. An additional low-speed centrifugation of 200 × g for 1 min was performed, separating the fixed cells of WB culture into a pellet and a supernatant fraction. The additional low-speed centrifugation enriched metaphase frequency and provided an MI comparable to the PBMC culture in the pellet fraction. Our study suggests that it is possible to increase the number of metaphase cells on slides using the slow centrifugation method, which could contribute to the efficiency of chromosome aberration analysis in biodosimetry.