Decoding Infection and Transmission · Published 2025-01-01 · DOI 10.1016/j.dcit.2025.100063
Objective: To establish and systematically characterize a contemporary cell line derived from Culex. tritaeniorhynchus collected in Hubei Province, China, with the goal of providing a standardized in vitro tool for investigations into mosquito biology and the molecular interactions between mosquitoes and mosquito-borne viruses (MBVs). Methods: Primary cell cultures were initiated from embryonic tissues of Cx. tritaeniorhynchus. Following continuous subculture for more than 50 passages, cell morphology and growth kinetics were recorded. Species identity was verified by sequencing of the mitochondrial cytochrome oxidase I (COI) gene. Susceptibility to eight arboviruses and two arthropod-specific viruses (ASVs) was evaluated using qRT-PCR, plaque assay and immunofluorescent staining. Transfection efficiency for eGFP expression was evaluated by fluorescence microscopy. Results: A stable cell line, designated CtrE1, was successfully established. The cells displayed uniform spindle-shaped morphology and maintained exponential growth (doubling time ranging from approximately 31 to 43 h). Molecular authentication confirmed Cx. tritaeniorhynchus origin. CtrE1 supported robust replication of six arboviruses and two arthropod-specific viruses, with genome copies reaching ≥107 copies μL-1 for Japanese encephalitis virus (JEV), Tembusu virus (TMUV) and Ebinur Lake virus (EBIV) within five days post-infection (dpi). Moderate transient eGFP expression was also achieved following plasmid transfection. Conclusion: CtrE1 was the first Cx. tritaeniorhynchus cell line derived from a contemporary Chinese population. Its broad virus susceptibility and genetic tractability provide a valuable in vitro platform for investigating mosquito–virus interactions, viral surveillance, and control strategies against JEV and other MBVs.
Abstract from DOAJ. Public domain (CC0 1.0).
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