The method of natural spawning is very passive and inconvenient for the study of developmental engineering in marine medaka, Oryzias dancena. The optimum concentration of human chorionic gonadotropin (HCG) and carp pituitary extract (CPE) for ovulation and spawning, and the injection time for the artificial spawning of marine medaka were analyzed in this study. The success rate, survival rate, and hatching rate were highest with 100 IU HCG kg-1 BW and 5 mg CPE L-1 in both male and female marine medaka (p<0.05). After obtaining unfertilized eggs and sperm by the injection of HCG and CPE into the broodstock of marine medaka, artificial fertilization could be successfully achieved any time fertilized eggs are needed in this species. This result should be useful for developing a study program for marine medaka as an experimental animal.