ESTABLISHMENT AND CHARACTERIZATION OF FIN CELL LINES FROM DIFFERENT PLOIDY ORIENTAL WEATHERFISH, MISGURNUS ANGUILLICAUDATUS

Oriental weatherfish (Misgurnus anguillicaudatus) is one of the most popular cultured species in China, Korean peninsula and Japanese archipelago. Primary culture of fin from diploid, triploid and tetraploid oriental weatherfish was performed with tissue explant method. Until now, cells of three tissues have been subcultured to passage 59 for diploid, 68 for triploid, and 68 for tetraploid, respectively. The special and combined aseptic processing method was immersing the fin tissue in 10% povidone-iodine for 15min, and then in penicillin and streptomycin mixture (500 IU/mL penicillin and 500 g/mL streptomycin) for 30min. The medium of primary culture and early subculture was DMEM/F12, supplementing with 20% fetal bovine serum (FBS), 10 ng/mL basic fibroblast growth factor(bFGF), 20 ng/mL insulin-like growth factor-I (IGF-I) and 10 g/mL chondroitin sulfate (pH7.2). The medium after passage 30 was only 20% FBS-DMEM/F12. The condition of cell culture was 25℃ with 5% CO2 continuously. The doubling time were 48.43h, 36.01h and 41.45h in diploid, triploid and tetraploid fin cells at the 50th passage, respectively. The feature chromosome number collected from 100 metaphase plates were 2n=50, 3n=75, 4n=100 in diploid, triploid and tetraploid fin cells, respectively. The cell livability of these three kinds of cell were (80.881.38)%, (84.481.13)% and (81.571.28)% when recovered after being stored in liquid nitrogen for 60d at the 40th passage. Establishment of fin cell lines from different ploidy oriental weatherfish in this study enriched the kinds of the fish cell lines, and would lay the foundation for discovering the mechanism of growth and genetics in polyploidy fish.