碩士 / 國立臺灣海洋大學 / 水產養殖學系 / 102 / Abstract
Sensitive cell line and its proliferation play important role in pathogenic viruses and preventative strategies. In this study, we cultured cell, derived from heart, bladder, kidney, spleen and embryo of four different spices grouper juveniles. There were 42 of developmental clones, but they couldn’t be survived over 40 passages. GE-1, a continuously new cell line was derived from orange spotted grouper (Epinephelus coioides ) embryo, was maintained in L-15 medium with 5% fetal bovine serum (FBS) at 25℃, and then it has been subcultured more than 100 times. The majority of GE-1 cells are epithelioid-like cells. Spontaneous transformation of GE-1 cells occurred during subculture from 35 to 65 of passages, and there was an increase in plating efficiency .The susceptibility of GE-1 cells to GIV (Grouper Iridovirus, Rana-type), GNNV (Grouper Nervous Necrosis Virus, nodavirus), TGIV (Grouper Iridovirus of Taiwan, Megalo-type) was observed Cytopathic effects (CPEs) 3-5 days post-infection, and the CPEs of TGIV was studied from 7 to 12 days post-infection. GIV was yielded titer of 108.8-9.3TCID50/ml. The titer of TGIV was decreased from 106.2TCID50/ml to 102.2 TCID50/ml. GNNV showed the same pattern like the titer of TGIV, the titer of the GNNV decrease from 105 to 102.8-2.9TCID50/ml. GH cells derived from orange spotted grouper heart was found in persistent infection with GIV by Nested PCR. GH supernatant cells was added into GE-1 cells, and then infected by GIV with different multiplicity of Infection (MOI). The result showed that added GH cells supernatant increased in survival rate after 24 hours with MOI=0.1,1. The conduction of Real-time PCR showed that the number of GIV copies were inhibited. Correlation of inhibition mechanism needed to be studied more further research.
| Identifer | oai:union.ndltd.org:TW/102NTOU5086015 |
| Date | January 2014 |
| Creators | Kang, Ren-Jie, 康人傑 |
| Contributors | Chou, Hsin-Yiu, 周信佑 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 56 |
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