Establishment and characterization of a cell line from the mosquito Culex tritaeniorhynchus (Diptera: Culicidae)

Abstract

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.

Argonaute 2 Suppresses Japanese Encephalitis Virus Infection in <i>Aedes aegypti</i>

“…Culture fluids were centrifuged (10,000 × g for 1 min) and then filtered through either a 0.45-or 0.22-mm Millex filter (EMD Millipore, Billerica, MA, USA) and titrated on Vero cells using a standard plaque assay (15,16). To count the number of plaques, Vero cells were fixed with 3.7z (v/v) formaldehyde in phosphate-buffered saline (PBS) for 1 h, the methylcellulose overlay was removed, and the cells were stained with methylene blue solution (16).…”

Mosquito cell lines: history, isolation, availability and application to assess the threat of arboviral transmission in the United Kingdom

“…GRIP-1Embryos[23]A.t. GRIP-2 & -3Larvae Aedes vexans West Nile virusUM-AVE1Embryos[26]Rift Valley fever virus Anopheles gambiae MalariaMos.55Larvae[27]Sua1BLarvae[28]4a-3BLarvae Anopheles stephensi MalariaMos.43Larvae[29]MSQ43Larvae[30] Anopheles albimanus MalariaLSB-AA695BBEmbryos[31] Culex quinquefasciatus West Nile virusUnnamedOvaries[32]Lymphatic filariasisUnnamedEmbryos[33] Culex theileri Rift Valley fever virusUnnamedEmbryos[34]Dog heartworm Culex tritaeniorhynchus Japanese encephalitis virusNIID-CTREmbryos[36] Culex bitaeniorhynchus Japanese encephalitis virusUnnamedEmbryos[37] Toxorhynchites amboinensis -TRA-171Larvae[38]…”

“…Cx. tritaeniorhynchus mosquitoes are the principal vectors of JEV in Asia, and cell lines have also been established from embryos of this species [35, 36]. The Cx.…”

“…The Cx. tritaeniorhynchus NIID-CTR cell line is highly susceptible to both JEV and DENV infection, and provides a valuable model for virus replication in the host [36]. Cx.…”

Characterization of Dak Nong virus, an insect nidovirus isolated from Culex mosquitoes in Vietnam

“…Coronaviruses and roniviruses, which have genomes larger than 26 kb, are categorized as ''large nidoviruses''; they are the largest RNA viruses known to date. In contrast, arteriviruses are referred to as ''small nidoviruses'' because of their [13][14][15][16] kb genome size.…”