Using Japanese Medaka Primary Hepatic Cell Culture to Investigate the Effect of Wastewater on Expression of VTG

• Main Authors: Yu-Ching Su, 蘇于菁
• Other Authors: Chien-Min Chen
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/96165685744381733705

碩士 === 嘉南藥理科技大學 === 環境工程與科學系碩士班 === 93 === In the last decade, much of savant efforts have been on studying endocrine-disrupting chemicals (EDCs), or commonly known as environmental hormones. These chemicals can imitate natural hormones and interfere not only the normal endocrinal, but also immune, reproductive and nerve systems in animals and humans. Some EDCs can be found in different water bodies or even in sewage-treatment works (STWs) effluents. The aims of this study were to investigate the effects of effluent from different sources on induction of vitellogenin (VTG) in the primary hepatocyte culture of Japanese medaka (Oryzias latipes) Because aquatic toxicology studies need to sacrifice a lot of animals, using cell culture can minimize the number of the animals used for different purposes. The medaka hepatocytes were cultivated in flasks with medium, and were exposed to 17-β estradiol and samples extracted from wastewater for 5days, 10days and 15days. The culture mediums were changed daily. VTG induction in hepatocytes was analyzed using Western blotting and quantified by the Bio-Rad’s image analysis system. The results showed that VTG induction by 17-β estradiol was dose-dependent after the 15-day exposure. Most of the influent samples from constructed wetlands could not induce vitellogenesis in cultured cells until exposed for 15 days. All of the effluent samples tested showed no VTG inducibility except those from An-Ping STW, and the responses were dependent with exposure time. Based on the results in this study, using Japanese medaka primary hepatic cell culture to investigate the effects of wastewater on induction of VTG was feasible, and at the same time, can be served as a alternative for in vivo study.