Trypanosoma cruzi: growth of clones on solid medium using culture and blood forms

Gomes, MA; Araújo, Silvana Márques de; Chiari, Égler

doi:10.1590/s0074-02761991000100022

Cited by 12 publications

(11 citation statements)

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“…Even when seeded on the same plate, trypanosomatids formed isolated colonies, each with a specific molecular karyotype. As mentioned previously, other studies have assessed the growth of T. cruzi on BLAB (2) or double-layered (3) media, resulting in plating efficiencies comparable to those in this study. Thus, depending on the aim of the study, SSL-HU can be used as an alternative to these other media.…”

supporting

confidence: 60%

“…In vitro cultivation of T. cruzi is used for detection of the parasite in vertebrate hosts and vectors. T. cruzi cultivation can be performed in liquid, solid, or semisolid media (2) (3) . Liver infusion tryptose (LIT) is a liquid medium (4) used for large-scale cultivation of trypanosomatids for genetic characterization and manipulation of parasites.…”

mentioning

confidence: 99%

“…In addition, growth differences have been observed between T. cruzi and T. rangeli on solid media (3) . One study showed that BLAB (BHI-LIT-blood agar), a solid medium (2) , resulted in the growth of T. cruzi colonies with reasonable plating efficiencies. However, growth of T. rangeli has not been assessed on BLAB medium.…”

mentioning

confidence: 99%

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Semisolid liver infusion tryptose supplemented with human urine allows growth and isolation of Trypanosoma cruzi and Trypanosoma rangeli clonal lineages

Fajardo

Cabrine-Santos

Ferreira

et al. 2016

Rev. Soc. Bras. Med. Trop.

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Introduction: This work shows that 3% (v/v) human urine (HU) in semisolid Liver Infusion Tryptose (SSL) medium favors the growth of Trypanosoma cruzi and T. rangeli. Methods: Parasites were plated as individual or mixed strains on SSL medium and on SSL medium with 3% human urine (SSL-HU). Isolate DNA was analyzed using polymerase chain reaction (PCR) and pulsed-field gel electrophoresis (PFGE). Results: SSL-HU medium improved clone isolation. PCR revealed that T. cruzi strains predominate on mixed-strain plates. PFGE confirmed that isolated parasites share the same molecular karyotype as parental cell lines. Conclusions: SSL-HU medium constitutes a novel tool for obtaining T. cruzi and T. rangeli clonal lineages.

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supporting

confidence: 60%

mentioning

confidence: 99%

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Semisolid liver infusion tryptose supplemented with human urine allows growth and isolation of Trypanosoma cruzi and Trypanosoma rangeli clonal lineages

Fajardo

Cabrine-Santos

Ferreira

et al. 2016

Rev. Soc. Bras. Med. Trop.

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“…Clones from the PH8 strain of L. amazonensis were obtained by plating in solid medium as described previously (Gomes et al, 1991;Tanuri et al, 1985) except for the modification of the culture medium (1% agar in alpha-MEM). Each cloned population was re-cloned three times.…”

Section: Leishmaniamentioning

confidence: 99%

The influence of ecto-nucleotidases on Leishmania amazonensis infection and immune response in C57B/6 mice

Souza¹,

Assis²,

Gomes³

et al. 2010

Acta Tropica

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Previous results from our laboratory and from the literature have implicated the expression of ecto-nucleotidases in the establishment of Leishmania infection. In the present study we evaluated the correlation between ecto-nucleotidasic activity and the infectivity of L. amazonensis promastigotes that were kept in culture for short or extended numbers of passages, a condition that is known to decrease parasite infectivity. We also analyzed the immune response associated with the infection by these parasites. As expected, we found that long-term cultured parasites induce the development of smaller lesions than the short-term cultured counterparts. Interestingly, long-term cultured parasites presented reduced ecto-nucleotidasic activity. In addition, cells recovered from animals infected with long-term cultured parasites produced higher amounts of IFN-gamma and have smaller parasite load, after 8weeks of infection. Furthermore, after 1week of infection, there is increased expression of the chemokine CCL2 mRNA in animals infected with short-term cultured parasites. Finally, infection of peritoneal macrophages by these parasites also shows marked differences. Thus, while short-term cultured parasites are able to infect a greater proportion of macrophages, cells infected by long-term cultured parasites express higher amounts of CXCL10 mRNA, which may activate these cells to kill the parasites. We suggest that the enzymes involved in metabolism of extracellular nucleotides may have an important role in infection by L. amazonensis, by acting directly in its adhesion to target cells and by modulating host cell chemokine production.

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“…Isolated clones from parasites transfected with pROCKPolhNeo were obtained as previously described by Gomes et al [1991]. Different numbers of parasites (10 2 , 10 3 , and 10 4 ) diluted in 0.1 mL of LIT medium were spread over an agar surface with a sterile glass loop and incubated at 288C.…”

Section: Selection Of T Cruzi Clonesmentioning

confidence: 99%

Cloning and characterization of DNA polymerase η from Trypanosoma cruzi: Roles for translesion bypass of oxidative damage

Moura

Schamber-Reis

Silva

et al. 2009

Environ and Mol Mutagen

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We report the cloning and characterization of the DNA polymerase eta gene from Trypanosoma cruzi (TcPoleta), the causative agent of Chagas disease. This protein, which can bypass cyclobutane pyrimidine dimers, contains motifs that are conserved between Y family polymerases. In vitro assays showed that the recombinant protein is capable of synthesizing DNA in undamaged primer-templates. Intriguingly, T. cruzi overexpressing TcPoleta does not increase its resistance to UV-light (with or without caffeine) or cisplatin, despite the ability of the protein to enhance UV resistance in a RAD30 mutant of Saccharomyces cerevisiae. Parasites overexpressing TcPoleta are also unable to restore growth after treatment with zeocin or gamma irradiation. T. cruzi overexpressing TcPoleta are more resistant to treatment with hydrogen peroxide (H(2)O(2)) compared to nontransfected cells. The observed H(2)O(2) resistance could be associated with its ability to bypass 8-oxoguanine lesions in vitro. The results presented here suggest that TcPoleta is able to bypass UV and oxidative lesions. However the overexpression of the gene only interferes in response to oxidative lesions, possibly due to the presence of these lesions during the S phase.

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Trypanosoma cruzi: growth of clones on solid medium using culture and blood forms

Cited by 12 publications

References 0 publications

Semisolid liver infusion tryptose supplemented with human urine allows growth and isolation of Trypanosoma cruzi and Trypanosoma rangeli clonal lineages

Semisolid liver infusion tryptose supplemented with human urine allows growth and isolation of Trypanosoma cruzi and Trypanosoma rangeli clonal lineages

The influence of ecto-nucleotidases on Leishmania amazonensis infection and immune response in C57B/6 mice

Cloning and characterization of DNA polymerase η from Trypanosoma cruzi: Roles for translesion bypass of oxidative damage

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