建立从杂交鳜（鳜鱼×斑鳜）脑细胞系中检测罗非鱼湖病毒。
Establishment of a brain cell line obtained from hybrids of Channa argus ×Channa maculata for the detection of tilapia lake virus.
机构信息
Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Liwan District, 510380, Guangzhou, Guangdong, China; College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 201306, China.
College of Animal Sciences and Technology, Zhongkai University of Agriculture and Engineering, Guangdong, Guangzhou, 510225, China.
出版信息
Microb Pathog. 2020 Jan;138:103810. doi: 10.1016/j.micpath.2019.103810. Epub 2019 Oct 22.
A brain cell line (CAMB) derived from hybrid snakehead (Channa argus (♂) × Channa maculata (♀)) was established by trypsin and collagenase combined digestion. The culturing conditions and cell biological characteristics were systematically studied. For growth of the cells, M199 medium containing 10% fetal bovine serum was used and at 27 °C incubated. Based on morphological analysis, CAMB cells were confirmed to be epithelial. The cell line has been subcultured more than 80 times since its initial primary culture. Chromosome analysis revealed that CAMB cells had an abnormal chromosome number 2n = 64, whereas the chromosome number in the hybrid snakehead was 45. The suitability of CAMB for tilapia lake virus (TiLV) was demonstrated. A CPE was observed after infection with TiLV-2017A. The highest TiLV titer was observed after 12 days post infection (dpi) and reached 107.2 TCID50/mL. The virus replication was confirmed by electron microscopic observations. Additionally, immunofluorescence assay confirmed the presence of TiLV-2017A after infection of CAMB. Therefore, CAMB cells can be a useful tool for the investigation of the pathogenesis of the TiLV induced disease in tilapia.
一个由杂交鳢（♂）×乌鳢（♀）衍生而来的脑细胞系（CAMB），通过胰蛋白酶和胶原酶联合消化法建立。对其培养条件和细胞生物学特性进行了系统研究。为了细胞生长，使用含 10%胎牛血清的 M199 培养基，在 27°C 下孵育。基于形态学分析，CAMB 细胞被确认为上皮细胞。自初始原代培养以来，该细胞系已传代培养超过 80 次。染色体分析显示，CAMB 细胞的异常染色体数为 2n=64，而杂交鳢的染色体数为 45。CAMB 适合感染罗非鱼湖病毒（TiLV）。感染 TiLV-2017A 后出现 CPE。感染后 12 天（dpi）达到最高 TiLV 滴度，达到 107.2 TCID50/mL。通过电子显微镜观察证实了病毒复制。此外，免疫荧光检测证实了 CAMB 感染后 TiLV-2017A 的存在。因此，CAMB 细胞可以作为研究罗非鱼 TiLV 诱导疾病发病机制的有用工具。
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