Tissue Culture from a Deep Sea Bivalve Calyptogena
Sumihiro KOYAMA*a, Akira INOUEa, and Masuo AIZAWAb
a The DEEPSTAR group, Japan Marine Science and Technology Center, 2-15 Natsushima- cho, Yokosuka 237, Japan
b Department of Bioengineering, Faculty of Bioscience and Biotechnology, Tokyo institute of Technology, 4259, Nagatsuta, Midori-ku, Yokohama 226, Japan
Studies of the biochemistry and molecular biology of deep-sea multicellular organisms has been hampered by the lack of established tissue Culture methods. In order to establish a novel tissue culture method for deep-sea organisms we have used the bivalve Calyptogena which was collected at a depth of 1180m by the unmanned submersible "Dolphin 3K". For a few days we were able to keep Calyptogena alive at atmospheric pressure.
The mantle tissue was cultured in DMEM medium prepared with artificial sea water supplemented with 10% FBS and 10% blood like body fluid of Calyptogena. The cells did not attach to the surface of non-coated dishes. If coated with poly- lysine, a majority of the cells stayed viable for at least 2 weeks and maintained a spherical shape.
The cell viability was estimated after staining with EthD-1 and calcein-AM to be around 90%. We were able to observe a slow increase in cell numbers supported by the appearance of mitotic cells.
In conclusion, we have indicated that the mantle tissue of the deep sea bivalve Calyptogena was viable in culture for 2 weeks, at 4℃ and atmospheric pressure.
