The use of an automated hematology analyzer to observe cell growth in the chromosome aberration test using human lymphocytes

Abstract

-Human lymphocytes have been frequently used for in vitro chromosome aberration or micronucleus tests on whole-blood culture. However, it is difficult to observe or confirm the cell growth of lymphocytes just before chemical treatment compared with cultured cell lines, such as CHL or CHO cells. In order to overcome this drawback of using whole-blood culture, we investigated a possibility of using an automated hematology analyzer (AHA) (Sysmex XT-2000i, SYSMEX Corp. (Hyogo, Japan)) to measure the growth of lymphocytes applying a manual function of this apparatus. In this study, wholeblood samples were cultured for 4 days, and the growth of lymphocytes was measured once a day using a standard flow cytometer (FCM) with antibody CD3 and DNA staining solution, and by the AHA simultaneously. The results showed that growth curves produced employing the two methods coincided fairly well. Therefore, it can be concluded that the growth of lymphocytes in whole-blood culture can be measured using AHA in a straightforward and rapid way in in vitro chromosome aberration or micronucleus tests.