Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium Isolamento e crescimento de Leishmania (Leishmania) infantum chagasi em meio de crescimento condicionado de cultura celular

• Published in: Revista do Instituto de Medicina Tropical de São Paulo
• Main Authors: Yeda L. Nogueira, Eunice A.B. Galati
• Format: Article in Journal/Newspaper
• Language: English
• Published: Universidade de São Paulo (USP) 2006
• Subjects: Leishmania (Leishmania) infantum chagasi; Flagellate form; Growth; McCoy Cell Lineage; Arctic medicine. Tropical medicine; RC955-962; Infectious and parasitic diseases; RC109-216; Arctic; McCoy
• Online Access: https://doi.org/10.1590/S0036-46652006000300006; https://doaj.org/article/c956659f8d964500abf3e92f147a1643

Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM). Leishmania (Leishmania) infantum chagasi (100 parasites) grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells) and its multiplication was observed for five days (120 hours). After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes). The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 µ L and photomicrographied. So the number of Leishmanias was adjusted to 1 mL volume. The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10² promastigotes) used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times) as observed by growth kinetics, within a shorter time. These results justify the use of this low-cost technique for the isolation and investigation of the behavior and multiplication of Leishmania both in vertebrates and invertebrates, besides offering means of obtaining antigens, whether whole antigens (leishmanias) or the soluble antigens produced by the parasites which may be useful for the production of new diagnostic kits. Leishmânias podem ser produzidas em meio de crescimento condicionado, após o cultivo de células McCoy (MCC). Promastigotas crescidas em meio semi-sólido NNN foram inoculadas em MCC, inicialmente, 100 parasitos por poço com 2,5 mL de McCoyMCC, em placas com 24 poços, sua multiplicação foi observada por uma cinética de 120 horas. Após este tempo, o meio estava saturado de promastigotas. A reprodução das leishmânias foi acompanhada a cada 24 horas, com contagem do número de parasitos por campo fotomicrografado. Como vantagem da técnica do crescimento da leishmânia em MCC tem-se o seu baixo custo, com pequena quantidade de parasitos pode-se obter o aumento da ...