Cultivos celulares primarios de Lutzomyia shannoni (Diptera: psychodidae) y estudio cariologico preliminar de la especie

Abstract

To obtain a Lutzomyia shannoni (Dyar) cell line for viral susceptibility studies and parasite maintainance, primary cell cultures of this species were initiated. This species is a vector for vesicular stomatitis virus in the United States and a suspected vector of cutaneous leishmaniasis in the Americas. Starting with embryos and phlebotominae neonate larvae, embryonic tissue explants in a MMIVP12 medium were carried out. After being supplemented with 20% fetal bovine serum and an antibiotic and antimycotic mixture, they were incubated at 28°C in a non-CO, environment. Cell growth began 85 to 88 days after the cultures were initiated dueto epithelial cell vesicle presence; these were floating in the media or adhered to small fragments of tissue with divided cells. Following mechanical stimulation of the cultures, cell proliferation increased in the week following the procedure; however, cell mitotic process was slow, similar to that developed with Lu. longipalpis, but different to cell cultures derived from mosquitoes. The formation of individual colonies, dispersed on the surface of the culture flasks, was observed 90 days after incubation; these later evolved to a semiconfluent monolayer. The cellular morphology was heterogeneous with predominant epithelial types. Using the squash technique, this species' karyotype was obtained; the number of diploid chromosomes was 8, derived from fourth-instar larvae brain tissue.