In Vitro Cultivation and Cryopreservation of

Leishmania Tropica Promastigotes

Abstract

Leishmaniasis is a parasitic disease caused by protozoans of the genus Leishmania and is a member of the group of neglected diseases. The lack of an effective vaccine and the increasing resistance of the parasites to drugs and of the vectors to insecticides are causing the disease to spread worldwide and remain a serious public health problem in many countries. In recent years, the significant weakening of entomological measures in the endemic regions of our country has led to outbreaks of leishmaniasis in various areas, posing serious epidemiological threats. Although serious measures were taken in our country before the 1990s, the epidemiological situation has significantly worsened over the last 30 years, and the disease has begun to appear in numerous regions of the republic. To date, the species of Leishmania parasites found in the individual endemic regions have not been identified or molecularly typed. Additionally, because isolation of the parasites has not been possible, the susceptibilities of the drug preparations used against them have also not been determined. To solve the aforementioned problems, the isolation of Leishmania parasites, the establishment of their continuous culture, and the creation of a cryobank are of paramount importance. This research article presents in detail the methodological approaches applied for the in vitro cultivation, cryopreservation, and establishment of a cryobank for Leishmania parasites. These studies, conducted at the Department of Vaccine and Nanovaccine Research of the V. Y. Akhundov Scientific Research Institute of Medical Preventive Medicine, provided information on the isolation of parasites, the establishment of continuous cultures, and the creation of a cryobank.

Keywords: Leishmania tropica, leishmaniasis, non-cultivation, biobank, cryopreservation

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