Thein VitroRadiosensitivity of Lymphocytes from Chronic Lymphocytic Leukaemia Using the Differential Staining Cytotoxicity (DiSC) Assay. I—Investigation of the Method

Abstract

A tumour sensitivity assay, the differential staining cytotoxicity (DiSC) assay, which has been shown to have potential in predicting tumour response to cytotoxic drugs, has been developed to investigate the radiosensitivity of peripheral blood lymphocytes isolated from patients with chronic lymphocytic leukaemia (CLL). The method involved irradiating isolated lymphocytes (0.64-20.48 Gy at 8 x 10(5) cells/ml) and incubating for 4 days. Subsequent radiation-induced cell kill was assessed by differential staining of dead and live cells and calculation of tumour cell viability. Factors affecting assay success rate and in vitro radioresponse were investigated. Results were shown to be reproducible both when repeat assays were performed on the same specimen on the same day, and on different specimens from the same patient after intervening periods of time. Greater than 1 day transit time (whole blood) or storage (separated cells in medium) was found to be detrimental to assay success, but pH of the medium (either pH 7.3 or 8) had little effect. An increased incubation period (from 4 to 7 days) slightly reduced cell survival, but the underlying radioresponsiveness of the cells did not change.