Infection, growth and maintenance of Wolbachia pipientis in clonal and non-clonal Aedes albopictus cell cultures.
Infection, growth and maintenance of Wolbachia pipientis in clonal and non-clonal Aedes albopictus cell cultures.
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克隆和非克隆白纹伊蚊细胞培养物中沃尔巴克氏菌的感染、生长和维持。
DOI：
10.1017/s0007485312000648
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发表时间：
2013
影响因子：
1.9
通讯作者：
Dobson,SL
中科院分区：
文献类型：
--
作者：
Khoo,CCH;Venard,CMP;Fu,Y;Mercer,DR;Dobson,SL
昆虫细胞系为研究生物系统提供了有用的体外模型，包括蚊子和专性细胞内共生体如Wolbachia pipientis之间的相互作用。白纹伊蚊Aa 23细胞系是第一个开发用于检测沃尔巴克氏体感染的细胞系。然而，使用Aa 23的沃尔巴克氏体研究可能因细胞系中不同细胞类型的存在和感染水平的实质性时间变化而复杂化。研究了两种方法来改善感染变异性。在第一种方法中，测试了多种Aa 23传代方案对感染变异性的影响。荧光原位杂交（FISH）染色用于表征随时间的沃尔巴克氏体感染水平。结果表明传代方法对沃尔巴克氏体感染水平有影响，某些方法导致感染丧失。所有传代方法均未成功有效缓解感染水平变化。在第二种方法中，克隆C7-10 A.用来自Aa 23细胞的沃尔巴克氏体和模拟果蝇（滨江）感染白纹伊蚊细胞系，分别产生命名为C7- 10 B和C7- 10 R的细胞系。通过FISH染色的表征显示相对于C7- 10 B中的感染，C7 - 10 R中的沃尔巴克氏体感染具有更大的稳定性和均匀性。Aa 23，C7- 10 B和C7- 10 R线的特性进行了讨论，作为研究沃尔巴克氏体宿主细胞相互作用的工具。
Insect cell lines provide useful in vitro models for studying biological systems, including interactions between mosquitoes and obligate intracellular endosymbionts such as Wolbachia pipientis. The Aedes albopictus Aa23 cell line was the first cell line developed to allow examination of Wolbachia infections. However, Wolbachia studies using Aa23 can be complicated by the presence of different cell types in the cell line and the substantial temporal variation in infection level. Two approaches were examined to ameliorate infection variability. In the first approach, multiple Aa23 passaging regimes were tested for an effect on infection variability. Fluorescence in situ hybridization (FISH) staining was used to characterize Wolbachia infection level over time. The results demonstrate an impact of passaging method on Wolbachia infection level, with some methods resulting in loss of infection. None of the passaging methods succeeded in effectively mitigating infection level variation. In a second approach, the clonal C7-10 A. albopictus cell line was infected with Wolbachia from Aa23 cells and Drosophila simulans (Riverside), resulting in cell lines designated C7-10B and C7-10R, respectively. Characterization via FISH staining showed greater stability and uniformity of Wolbachia infection in C7-10R relative to the infection in C7-10B. Characterization of the Aa23, C7-10B and C7-10R lines is discussed as a tool for the study of Wolbachia-host cell interactions.