Inhalant anesthesia is routinely used for cesarian section in many animal species, allowing the safe delivery of neonates and smooth recovery of dams. However, in mice, inhalant anesthesia in cesarean section may be avoided due to fear of negative health effects on retrieved pups. This study compared the effects of isoflurane anesthesia on pups after cervical dislocation of conscious and anesthetized dams. Time-mated C57BL/6J dams were either anesthetized with 5% isoflurane or were conscious during cervical dislocation. Rederived pups were fostered to Swiss Webster dams and weaned at 21 d. Weights of litters were recorded at birth, and individual pup weights were recorded at weaning. We found no significant difference between the two treatment groups in pup survival until weaning. We also found no significant difference when comparing the average weaning weights of all the male pups to that of all the female. Female pups from isoflurane-treated dams had significantly higher weaning weights than did those from unanesthetized dams; however, the weights of male pups from the two groups were not different at weaning. This study found no immediate negative effects of using isoflurane anesthesia prior to cervical dislocation of C57BL/6J pregnant dams for the purpose of rederivation. Isoflurane can be used for cervical dislocation of pregnant C57BL/6J dams without affecting pup survival.