Primary Psorophora confinnis (Dipthera: Culicidae) cell culture studies

Abstract

With the purpose of obtaining a Ps. confinniscell line for arbovirus infection susceptibility studies, the primary cultures of this species were initiated, this being a vector of the Venezuelan Equine Encephalitis virus, epidemo-epizootic type. Starting from embryonated eggs, neonate larvae and adult females' ovaries, embryonic tissue explants in diverse culture media (supplemented with 20% foetal bovine serum and a mixture of antibiotics and 1% antimycotics) were carried out. Biological material was sterilised by immersion in different substances, such as 1.6% sodium hypochloride, 70% ethanol and 0.25% mercuric chloride solution in 70% ethanol. Cellular growth only began after 62 days of incubation in MMNP12 medium; proliferation of isolated colonies also started from larval fragments' cut ends. Evolution of cellular growth until confluent monolayer formation was extremely slow and was only reached 8 months post-explant, presenting a predominantly epithelioid cellular morphology. Cell growth was not successful starting from adult female ovarian tissue. lnitiation of this specie's cell growth presented different times compared to that spent on cell cultivation of other mosquitoes, which indicates that, in spite of using similar methodology in the process in order to obtain primary cell cultures, cell adaptations to physical, environmental and nutritional conditions are different for each one of the,species. This is the first report of cell cultures of a species of mosquito belonging to the Psorophora genus.