Ovarian primary tissue culture of the kuruma shrimp Marsupenaeus japonicus

Cell growth in ovarian primary culture of the kuruma shrimp, Marsupenaeus japonicus, was examined under various culture conditions. The best growth of ovarian cells was obtained in a culture system consisting of double strength of Leibovitz-15 medium supplemented with 10% fetal bovine serum, glucose (1 g/L), proline (0.1 g/L), TC-Yeastolate (1 g/L), and lactalbumin hydrolysate (1 g/L). The cells survived in this medium at 25 degrees C for 45 d. The epithelial-like cells predominated in 10-d-old cultures, covering >80% of the surface area on the bottom of flask. Cells in mitosis were often observed. Cell proliferation was monitored by incorporation of 5-bromo-2'-deoxyuridine (BrdU), an analog of thymidine. 5-Bromo-2'-deoxyuridine-associated cells accounted for 11.5 and 35.0% of cell populations at 2 and 24 h, respectively, after BrdU treatment. Our results provide an improved culture technique for ovarian tissue of the kuruma shrimp.