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Establishment and characterization of a cell line from Ghanaian Aedes aegypti (Diptera: Culicidae) focusing on Aedes -borne flavivirus susceptibility
In Vitro Cellular & Developmental Biology - Animal ( IF 1.5 ) Pub Date : 2020-09-30 , DOI: 10.1007/s11626-020-00504-y
Michael Amoa-Bosompem 1, 2, 3 , Daisuke Kobayashi 1 , Kentaro Itokawa 4 , Astri Nur Faizah 1, 5 , Ryusei Kuwata 6 , Samuel Dadzie 3 , Takaya Hayashi 7 , Shoji Yamaoka 7 , Kyoko Sawabe 1 , Shiroh Iwanaga 2 , Haruhiko Isawa 1
Affiliation  

Mosquitoes are generally considered one of the most important vectors of arboviruses, with Aedes aegypti regarded as the most important in transmission of yellow fever and dengue viruses. To investigate why there are differences in the incidence of dengue fever and Zika in different geographical areas and an absence of outbreaks in Ghana in spite of an abundance of A. aegypti mosquitoes, we established a continuous cell line from embryonic cells of A. aegypti collected in Ghana and assessed its susceptibility to dengue, yellow fever, and Zika viruses. The new cell line (designated AeAe-GH98), having an adhesive spindle-shaped web-like morphology, was serially subcultured in both VP-12 and Schneider’s medium supplemented with 10% heat-inactivated fetal bovine serum. AeAe-GH98 cells were found to have a population doubling time of 1.3 d during exponential growth. The mosquito colony used to establish the cell line was confirmed to have originated from Africa using microsatellite assay. In terms of susceptibility to Aedes-borne flaviviruses, AeAe-GH98 cells were found to have different degrees of susceptibility to yellow fever, Zika, and dengue virus infection and propagation. While susceptibility of AeAe-GH98 cells to yellow fever and Zika viruses was comparable with that of C6/36 cells, susceptibility to dengue virus was significantly lower. This cell line will serve as a useful tool for determining molecular factors influencing virus–vector susceptibility in vitro.



中文翻译:

以伊蚊传播的黄病毒易感性为重点的加纳伊蚊(双翅目:库利科)细胞系的建立和表征

蚊子通常被认为是虫媒病毒最重要的载体之一,埃及伊蚊被认为是传播黄热病和登革热病毒的最重要媒介。为了研究为什么不同地理区域的登革热和寨卡病毒的发病率存在差异,并且尽管有大量埃及蚊,但加纳没有爆发疫情,我们从埃及埃及蚊的胚胎细胞中建立了一个连续的细胞系在加纳收集并评估其对登革热、黄热病和寨卡病毒的易感性。新细胞系(命名为 AeAe-GH98)具有粘性纺锤形网状形态,在 VP-12 和施耐德培养基中连续传代培养,并辅以 10% 热灭活胎牛血清。发现 AeAe-GH98 细胞在指数生长期间具有 1.3 天的群体倍增时间。使用微卫星分析证实用于建立细胞系的蚊子群起源于非洲。在对伊蚊的易感性方面经发现,AeAe-GH98 细胞对黄热病、寨卡病毒和登革热病毒感染和繁殖具有不同程度的易感性。虽然 AeAe-GH98 细胞对黄热病和寨卡病毒的易感性与 C6/36 细胞相当,但对登革热病毒的易感性明显较低。该细胞系将作为确定影响体外病毒-载体敏感性的分子因素的有用工具。

更新日期:2020-10-02
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