Apoptosis of isolated rat splenocytes induced by γ-irradiation

Objective To investigate apoptosis of splenocytes induced by γ-irradiation in vitro.Methods The spleens were excised from Wistar rats under aseptic condition,and single-cell suspension of splenocytes was prepared.After incubation in RPMI 1640 medium supplemented with 10% fetal calf serum for 24～48 hours,splenocytes were irradiated by γ rays at different dosages and time courses.The morphological changes were observed,and apoptotic rates of splenocytes were detected by flow cytometry and DNA Ladder determination.Results After γ-irradiation,splenocytes demonstrated the characteristic morphological and molecular biological changes.Flow cytometry assay showed that 5 000 rad and 10 000 rad γ-irradiation induced 76.63%±3.53% and 82.99%±2.04% of apoptotic rates,respectively;which were significantly different (P0.005) from that in the control group (36.04%±2.98%).Conclusion The γ-irradiation could efficiently induce splenocyte apoptosis in vitro,which may be used for further exploration of immune mechanism and clinical application.