Department of Medical Entomology, National Institute of Infectious Diseases, Tokyo, Japan.
In Vitro Cell Dev Biol Anim. 2012 Jun;48(6):369-76. doi: 10.1007/s11626-012-9520-1. Epub 2012 Jun 16.
We established a continuous cell line from the embryo of the mosquito Culex tritaeniorhynchus Giles (Diptera: Culicidae), a known major vector of the Japanese encephalitis virus (family Flaviviridae, genus Flavivirus) in Asia. The cell line, designated NIID-CTR, was serially subcultured in VP-12 medium supplemented with 10 % heat-inactivated fetal bovine serum (FBS). It continued to grow for more than 60 passages over a 750-d period. The NIID-CTR cell line mainly comprised two morphologically distinct types of cells with adhesive properties: spindle-shaped and round cells. Most of the NIID-CTR cells at the 45th passage were diploid (2n = 6). The growth kinetics of the NIID-CTR cells was significantly affected by the FBS concentration in the medium. The population doubling time of the NIID-CTR cells was 20 h in the presence of 10 % FBS and 76 h in its absence. The DNA sequence of the mitochondrial cytochrome oxidase I gene confirmed that the NIID-CTR cell line was derived from C. tritaeniorhynchus. The cells were highly susceptible to Japanese encephalitis and Dengue viruses, thus providing a valuable tool for the study of mosquito-borne flaviviruses.
我们从亚洲已知的日本脑炎病毒（黄病毒科，黄病毒属）主要媒介三带喙库蚊（双翅目：库蚊科）的胚胎中建立了一个连续细胞系。该细胞系命名为 NIID-CTR，在补充有 10%热灭活胎牛血清（FBS）的 VP-12 培养基中连续传代培养。在 750 天的时间里，它超过 60 代持续生长。NIID-CTR 细胞系主要由两种具有粘附特性的形态上明显不同的细胞类型组成：梭形和圆形细胞。在第 45 代时，大多数 NIID-CTR 细胞为二倍体（2n=6）。NIID-CTR 细胞的生长动力学受培养基中 FBS 浓度的显著影响。在存在 10%FBS 的情况下，NIID-CTR 细胞的倍增时间为 20 小时，而不存在 FBS 的情况下为 76 小时。线粒体细胞色素氧化酶 I 基因的 DNA 序列证实 NIID-CTR 细胞系源自三带喙库蚊。这些细胞对日本脑炎和登革热病毒高度易感，因此为研究蚊媒黄病毒提供了有价值的工具。
