Effect of Human Urine on Cell Cycle and Infectivity of Leismania Species Promastigotes In Vitro

Allahverdiyev, Adil M.; Bağırova, Melahat; Elcicek, Serhat; Çakır-Koç, Rabia; Oztel, Olga Nehir

doi:10.4269/ajtmh.2011.10-0207

Cited by 15 publications

(14 citation statements)

References 32 publications

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“…Human urine is a clear example of a media component with a specific biological imprint. When used as a medium supplement it increases parasite proliferation (Shamsuzzaman et al 1999) and infectivity (Allahverdiyev et al 2011).…”

Section: Introductionmentioning

confidence: 99%

The impact of distinct culture media inLeishmania infantumbiology and infectivity

et al. 2013

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An ideal culture medium for Leishmania promastigotes should retain the basic characteristics of promastigotes found in sandflies (morphology and infectivity). Furthermore, the media should not create a bias in experimental settings, thus enabling the proper extrapolation of results. To assess this we studied several established media for promastigote growth. We analysed morphology, viability, cell cycle progression, metacyclic profile, capacity to differentiate into axenic amastigotes and infectivity. Furthermore, using a rational approach from the evaluated media we developed a simple serum-free medium (cRPMI). We report that parasites growing in different media present different biological characteristics and distinct in vitro and in vivo infectivities. The developed medium, cRPMI, proved to be a less expensive substitute for traditional serum-supplemented media for the in vitro maintenance of promastigotes. In fact, cRPMI is ideal for the maintenance of parasites in the laboratory, diminishing the expected loss of virulence over time typical of the parasite cultivation. Ultimately this report is a clear warning that the normalization of culture media should be a real concern in the field as media-specific phenomena are sufficient to induce biological bias with consequences in infectivity and general parasite biology.

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Section: Introductionmentioning

confidence: 99%

The impact of distinct culture media inLeishmania infantumbiology and infectivity

et al. 2013

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“…Vitamins act as co-factors enabling many enzymes to function properly in cells, and in some cases are essential where cells cannot synthesize them, so a vitamin supplement is often required in media. Several reports have shown that 1-5% (v/v) human urine can stimulate the growth of Leishmania promastigotes ( Howard et al., 1991 ; Singh et al., 2000 ; Iqbal et al., 2006 ; Allahverdiyev et al., 2011 ). Allahverdiyev et al.…”

Section: Discussionmentioning

confidence: 99%

Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes

Chanmol

Jariyapan

Preativatanyou

et al. 2022

Front. Cell. Infect. Microbiol.

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Leishmania (Mundinia) orientalis is a human pathogen causing leishmaniasis and studies on the properties of metacyclic promastigotes, the parasite’s infective stage, are required for a better understanding of its transmission and infection. However, information on cultivation for mass production of L. orientalis metacyclic promastigotes and factors that stimulate their metacyclogenesis is limited. Therefore, the objective of this study was to develop a suitable methodology for generating promastigote cultures containing a high proportion and number of L. orientalis metacyclic promastigotes. Various media, i.e., Schneider’s insect medium, Medium 199 and Grace’s insect medium, supplemented with various quantities of dithiothreitol, Basal Medium Eagle vitamins, pooled human urine, and fetal bovine serum, were optimized for metacyclogenesis. The results revealed that the optimum culture medium and conditions of those tested were Schneider’s insect medium supplemented with 100 μM dithiothreitol, 1% (v/v) Basal Medium Eagle vitamins, 2% (v/v) pooled human urine, and 10% (v/v) fetal bovine serum, pH 5.0 at 26°C. We also demonstrated that L. orientalis metacyclic promastigotes could be purified and enriched by negative selection using peanut lectin. Under these culture conditions, the highest yield of metacyclic promastigotes was obtained with a significantly higher percentage of parasite survival, resistance to complement-mediated lysis, and infection index in THP-1 macrophage cells compared to parasites cultured without media supplements at neutral pH. This is the first report providing a reliable method for mass production of L. orientalis metacyclic promastigotes for in vivo infections and other experimental studies of this emerging parasite in the future.

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“…This study mentioned that higher concentrations of urine were not as good, probably because of a fall in pH and inhibitory concentrations of human metabolites [13]. But a research reported that human urine in parasite cultures differentially affected and increased the infectivity and proliferation of Leishmania species promastigotes and facilitated the transition from G0/G1 to S phases more quickly and their results have shown that promastigotes could efficiently proliferate and grow in culture media containing relatively high (25%) urine concentrations [40]. In our previous researches about finding suitable replacements for FCS we had found that the serum and the urine of many animals like hamster, rabbit, and sheep are suitable for cultivation of promastigotes, but, collecting the serum of some of them, like Hamster, is expensive and using of some of them, like sheep serum, is accompanied with many challenges about adaptation problems of promastigotes to new serum (unpublished work).…”

Section: Discussionmentioning

confidence: 99%

Effects of Sheep and Mouse Urine on the Growth Pattern ofLeishmania majorPromastigotes

Nasiri

Karimi

Dalimi

et al. 2013

BioMed Research International

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The protozoan parasites of the genus Leishmania are the causative agents of different clinical diseases. Fetal calf serum (FCS) is the main part and the most expensive ingredient of the Leishmania culture media. Here, the efficacies of different concentrations (1%, 2.5%, 5%, and 10%) of the filtered and autoclaved sheep and mouse urine were evaluated as a growth stimulator in Leishmania culture procedure. The results indicated that culture media enriched with the filtered sheep and mouse urine supported the growth of the parasites and can be used for cultivation of Leishmania parasites. In conclusion, this study has demonstrated an alternative low-cost medium that could be used in cultivation process of Leishmania major promastigotes.

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Effect of Human Urine on Cell Cycle and Infectivity of Leismania Species Promastigotes In Vitro

Cited by 15 publications

References 32 publications

The impact of distinct culture media inLeishmania infantumbiology and infectivity

The impact of distinct culture media inLeishmania infantumbiology and infectivity

Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes

Effects of Sheep and Mouse Urine on the Growth Pattern ofLeishmania majorPromastigotes

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