Primary cell culture for Taiwan abalone,Haliotis diversicolor

• Main Authors: Jo-Lan Huang, 黃若蘭
• Other Authors: Chang-Jen Huang
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/62361110416147012866

碩士 === 國立臺灣海洋大學 === 生物科技研究所 === 95 === Taiwan abalone (Haliotis diversicolor) is one of the important economical aquaculture shellfish in Taiwan. In 2002-2003, a massive death break out occurred in north-east of Taiwan, and virus was found to be the causative pathogen. The objective of this study is to establish tissue culture system of Taiwan abalone as the platform for studying virus infection in the future. In the preliminary data, the adhesion rate of Taiwan abalone cells was augmented by rising the concentration of L-15 and Grace’s medium. Therefore, the medium osmolarity is probably the key factor for Taiwan abalone cells adhesion. We also observed that high osmolarity (735~1,110 mOsmkg-1) of L-15 medium can promote abalone ovarian cells adhesion, and the proliferation can also be stimulated by the supplement of salt solution. In order to search better condition of abalone cell culture, we have developed a new medium MAH by modifying the formula of L-15 and Grace’s medium and also from the results of component assay of Taiwan abalone hemolymph extract. Comparing the growth of Taiwan abalone ovarian and hemocyte cells in modified LS20, GLS15 and MAH media, the best medium is GLS15. Although the adhesion cells are unable to attach after subculture, the suspension cells can be passaged. Till now, the suspension ovarian cells can be subcultured for 8 times, alive for 85 days; the suspension hemocyte can be subcultured for 11 times, alive for 96 days. In the medium supplement study, the supplement of abalone hemolymph extract can promote adhesion and proliferation of hemocyte, but not of insulin.