The study of the application of poultry hepatocytes in biodetection of environmental hormones.
| Autor: | Shiau-Wei Chen, 陳筱薇 |
|---|---|
| Rok vydání: | 2007 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 95 The purpose of this study was to utilze to-be-discarded layer cockerels to establish the poultry hepatocyte primary cell culture system and to illustrate the feasibility of using in detecting the environmental hormones. The hepatocytes were obtained from the liver tissue following perfusing and disassociation. Hepatocytes were treated with 17 β-estradiol (E2) for two days and prepared for mRNA isolation after culturing. Vitellogenin (VTG) gene expression was examined via reverse-transcription polymerase chain reaction (RT-PCR). In the poultry hepatocyte primary cell culture system, the yield of hepatocytes were about 107 cells per gram of perfused liver tissue, and the average viability was about 80%. After culturing, the hepatocytes attached to the culture dish and grow up normally. The viability rate was about 94%. Albumin experiment in hepatocytes was examined to ralidate the ascertainment of hepatocyte. In vivo experiment showed that the VTG gene expression was induced faster and stronger in pullets then in cockerels. If layer cockerels were not primed with E2 ahead of time, the VTG gene would not express normally in following injection. VTG gene expression was more observable after two days of inject. Results from in vitro experiment showed that VTG gene expressed was dose dependent when stimulated with 10-5~10-7 M E2. However when they are stimulated with 10-4M E2, VTG was not expressed. This result may be due to cell toxicity. According to the study, the hepatocyte cell culture system developed by to-be-discarded layer cockerel can be applied to detect environmental hormones via examining the E2-induced VTG gene expression. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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