The effect of mouse euthanasia technique on subsequent lymphocyte proliferation and cell mediated lympholysis assays

The purpose of this study was to determine the effects that specific euthanasia methods have on mitogen induced lymphocyte proliferation (LP) and the induction of alloantigen specific cytolytic T-lymphocytes (CTL). Mice were euthanatized by cervical dislocation (CD), or anesthesia with methoxyflurane or pentobarbital followed by CD (M-CD or P-CD respectively), CO2 overexposure (CO2-OD) or halothane overexposure (H-OD). Mitogenic lymphoproliferation was increased in cells derived from mice euthanatized by M-CD and P-CD. In contrast, the cytolytic profile of CTL derived from mice euthanatized by P-CD, CO2-OD and H-OD was decreased. The results of this study show that euthanasia techniques involving the use of methoxyflurane, pentobarbital, CO2 and halothane affect in vitro lymphoproliferation and CTL function. We conclude that the method of euthanasia influences certain immunologic parameters and selection of a particular technique should be given careful consideration.