In this study, we tried to maintain the vitality of Chinese sturgeon (Acipenser sinensis) ova before fertilization with several treatments in vitro. The ovulated eggs were allocated to groups with different incubation medium (coelomic fluid and artificial media), temperature (4°C and 16°C) and storage duration (2 h and 6 h). The maximum fertilization and hatching rate were observed for the control group in which the ova were fertilized immediately after spawning, with the values of 82.45% and 84.73%, respectively. Compared with the control group, the fertilization and hatching rate of all the treatment groups stored at 4°C or in coelomic fluid decreased significantly (P < 0.05). The fertilization rate of the treatment group stored in artificial medium at 16°C did not change obviously in the first 2 h (P > 0.05), but declined dramatically (P < 0.05) after 6 h. In comparison with the control group, no significant (P > 0.05) reduction was shown in hatching rate of the treatment group stored in artificial medium at 16°C for 6 h. The results showed that the ova of Chinese sturgeon can be stored for at least 6 h at 16°C in artificial medium without weakening; this provides a practical application method for the routine hatchery practice of Chinese sturgeon, as well as certain relevant research.