Chinese sturgeon (Acipenser sinensis) is an endangered species, listed as a Grade I protected animal in China. The females rarely successfully develop their gonads from stage II to III in captivity, which handicaps the propagation of cultured Chinese sturgeon. The current study aimed to understand the effects of dietary lipid level on the ovarian development and the related regulation mechanism in female Chinese sturgeon. A 24-month feeding trial was conducted with 10-year-old Chinese sturgeons with ovaries at the developmental stage II, with three experimental diets containing 10, 14 and 18% lipids. Ovary, muscle and serum samples were collected at four time-points (6, 12, 18 and 24 months) for further analyses. Serum metabolomics and ovary transcriptomics analyses were conducted at the 18 months. Results showed that only the 18% lipid diet promoted ovary development to the stage IV. Oocytes at stage II in this group also exhibited higher diameter and more lipid droplets. Serum triglyceride content in the 18% group was significantly higher than in groups 10 and 14% (both at 12 and 18 months). Estradiol content in the group 14% was significantly higher than in 10 and 18% groups, except at 24 months. Metabolomic and transcriptomic results indirectly indicated that 14% of dietary lipids benefited steroid hormone synthesis, while 18% lipid facilitated arachidonic acid metabolism, cholesterol biosynthesis and vitellogenesis, although serum cholesterol content did not vary with the dietary lipid level. In conclusion, 18% dietary lipid is the optimal level for improving gonad development of female Chinese sturgeon.