Ascidian hemocytes culture and the prevention of bactirail contamination
海鞘血细胞培养和细菌污染控制

In this study, ascidian hemocytes were cultured in vitro and the identification and prevention of bacterial contamination were investigated. After 1 h incubation, hemocytes spread and formed a flat sheet consisting of nine types of cells. Most cultured cells were round and several cells were amebocytes. Round cell types could be maintained for 2 weeks at least in vitro culture, whereas in vitro could survived for very short time. Two bacterial strains contaminated in cell culture were isolated from the bacterial contamination samples by culture and purity culture. Two bacterial strains were gram-negative bacterium. The detected bacteria were isolated and identified as the members of by 16S rDNA sequencing. Drug sensitivity test detected that Vibrio showed resistance to chloromycetin and andciprofloxacin, Shewanella showed resistance to imipenem and chloromycetin. In order to control the microbial contamination in cell culture, three antibiotics mixtures were compared. It is shown that gentanycin was efective to prevent bacterial contamination.