Effect of storage conditions of blood on radiation-induced chromosomal aberrations and apoptosis in human lymphocytes

Abstract

To evaluate the effect of storage conditions of blood on the direct relationship between radiation-induced chromosome aberrations and apoptosis in human peripheral blood lymphocytes,whole bloodwas irradi- atedwith 3Gy X-rays. Directly after irradiation, a sample of bloodwas analyzed for chromosome damage and proliferation index, after phytohaemagglutinin stimulation and incubation at 37 ◦ C for 56 h. Blood samples were stored for 48 h at 4 and 20 ◦ C with or without phytohaemagglutinin and analyzed for cell viability and apoptosis at 0, 24 and 48 h storage time. After 48 h of storage, unstimulated cultures were stimulated to proliferate. These samples and cultures stimulated immediately before storage were incubated at 37 ◦ C for 56 h and analyzed for chromosome damage and proliferation index. Metaphases were examined for the presence of dicentrics, excess acentrics, and rings. Storage at 20 ◦ Cwithout phyto- haemagglutinin for 48 h increases significantly the yield of apoptosis and decreases significantly the yield of dicentrics. During 48 h of storage time the presence of phytohaemagglutinin and the temperature of 4 ◦ C protected the irradiated lymphocytes from apoptosis allowing accurate estimation of the real yield of radiation-induced chromosome damage. Therefore these blood-storage conditions enable analysis in metaphase and may offer some advantages for biodosimetry of absorbed radiation dose.