Cultivation of Leishmania braziliensis in an Economical Serum-Free Medium Containing Human Urine

Abstract

Leishmania braziliensis cells are difficult to culture in vitro and usually require media supplemented with serum for sustained cell division. Fresh, sterile urine is an inexpensive substitute for serum in the culture of 2 strains of L. braziliensis, 1 infected with Leishmania RNA virus 1, and 1 uninfected. In the presence of urine, both the infected and the uninfected strains grew to the same final cell density as the same strains grown in the presence of serum. One strain of Leishmania major was also successfully cultured in urine-supplemented media.

Leishmania spp: completely defined medium without serum and macromolecules (CDM/LP) for the continuous in vitro cultivation of infective promastigote forms.

“…Other strains or species have been adapted to continuous growth in less rich media containing serum or with serum replaced by bovine serum albumin, 20,21 bacteriologic media, or more recently, human urine. 32,33 The complexity of the media used to cultivate members of the genus Leishmania has complicated the feasibility of comparative studies on nutritional requirements, metabolism, and other characteristics of these organisms, especially those isolated from mammals.…”

“…These media were enriched with large concentrations of certain amino acids, vitamins, bovine albumin, hormones, peptide supplements, [25][26][27][28][29][30][31] or more recently, with human urine. 32,33 In the present paper, we report a completely chemically defined culture medium free of serum, macromolecules, proteins, and peptides that readily supports the growth and maintenance of promastigote forms of various Leishmania species without compromising parasite growth rates. Morphologic, biochemical, immunologic, and biologic properties of aserically grown promastigote forms are presented.…”

LB broth-lyophilized Rabbit serum (LLR) as a new and suitable culture medium for cultivation of promastigotes of Leishmania major

“…The many of different kinds of media that have been using for cultivation of genus Leishmania require expensive and batch to batch different quality fetal calf serum (FCS), that is not manufactured in many countries specially most of the poor tropical countries, as one of their essential ingredients. FCS is highly expensive, and reliable supply is very difficult to obtain, especially in developing countries (Newman 2003) and using it in cell culture is problematic for several reasons as complexity, highly variable, and difficult to characterizing (Armstrong and Patterson 1994). Several attempts have been made to replace FCS in leishmania culture media with different kinds of sera, bovine serum albumin, a mixture of purine bases, vitamins, large concentrations of certain amino acids, hormones, hemin, hemoglobulin, human and animal urine and, more recently chicken serum (Trager 1953;Herman 1966;Ghoshal et al 1986;Ali et al 1998;Merlen et al 1999;Shamsuzzaman et al 1999;Pal and Joshi-Purandare 2001;Schuster et al 2002;Nasiri et al 2013a, b;Nasiri 2013) that the later introduced an alternative low-cost serum that can be used in culture medium for primary isolation, routine cultivation and mass cultivation of Leishmania parasites (Nasiri et al 2011(Nasiri et al , 2013a.…”

Estudo comparativo de técnicas de demonstração de amastigotas e isolamento de promastigotas no diagnóstico da leishmaniose tegumentar americana

“…5 Armstrong e colaboradores relataram que a urina humana estéril adicionada ao meio de cultura também estimula a divisão celular de formas promastigotas e facilita o cultivo primário de células infectadas de tecido animal, podendo substituir o soro fetal bovino nos meios de cultura. 6 Foram usadas, em ambos casos, cepas já identificadas e mantidas em criopreservação.…”

“…5 Armstrong e colaboradores relataram que a urina humana estéril adicionada ao meio de cultura também estimula a divisão celular de formas promastigotas e facilita o cultivo primário de células infectadas de tecido animal, podendo substituir o soro fetal bovino nos meios de cultura. 6 Foram usadas, em ambos casos, cepas já identificadas e mantidas em criopreservação.O objetivo deste estudo foi verificar se a urina humana estéril, adicionada ao meio Difco e Scheneider, seria mais eficiente ou poderia substituir o soro bovino fetal em cultivos de espécies de Leishmanias existentes no país, bem como se as técnicas de esfregaço, com coleta do material por biópsia ou matchstick, poderiam ter sensibilidades diferentes. …”