Establecimiento de un sistema in vitro para el cultivo de Leishmania chagasi/ infantum en macrófagos de perro.

Establishing an in vitro system for culturing Leishmania chagasi/infantum in dog macrophages Few in vitro studies for culturing Leishmania chagasi/infantum in dog macrophages have been published. The use of canine macrophages to obtain amastigotes allows the study of the biology of this parasitic form in the host cell. In order to determine the conditions suitable for the establishment of an in vitro system for culturing Leishmania chagasi/infantum amastigotes, the etiological agent of American Visceral Leishmaniasis, dog macrophages belonging to the continuous cell line DH82 were infected using promastigotes under different conditions. It was observed that, although cells ingested promastigotes very quickly, the parasites were transformed into amastigotes and the dog macrophages were able to withstand intracellular replication of the amastigotes; moreover. In the cellular growth phase, the parasite: cell ratio and the incubation temperature have an infl uence on the percentage of infected cells. The highest infection rates were obtained when infected monolayer cells were in a stationary phase, using 50:1 parasite:cell ratio, and incubating at 33oC, which suggests that these are the ideal conditions for developing the in vitro culture of these amastigotes. These fi ndings open the possibility of using this cell line in the study of parasite-host interactions, as well as evaluating drugs with a potential as a leishmanicide agent.