Ítem
Acceso Abierto

Purification of Trypanosoma cruzi metacyclic trypomastigotes by ion exchange chromatography in sepharose-DEAE, a novel methodology for host-pathogen interaction studies

Mostrar el registro sencillo de la publicación
dc.creatorCruz-Saavedra L.spa
dc.creatorMuñoz M.spa
dc.creatorLeón C.spa
dc.creatorPatarroyo M.A.spa
dc.creatorArevalo G.spa
dc.creatorPavia P.spa
dc.creatorVallejo G.spa
dc.creatorCarranza J.C.spa
dc.creatorRamírez, Juan Davidspa
dc.date.accessioned2020-05-25T23:56:21Z
dc.date.available2020-05-25T23:56:21Z
dc.date.created2017spa
dc.description.abstractMetacyclic trypomastigotes are essential for the understanding of the biology of Trypanosoma cruzi, the agent of Chagas disease. However, obtaining these biological stages in axenic medium is difficult. Techniques based on charge and density of the parasite during different stages have been implemented, without showing a high efficiency in the purification of metacyclic trypomastigotes. So far, there is no protocol implemented where sepharose-DEAE is used as a resin. Therefore, herein we tested its ability to purify metacyclic trypomastigotes in Liver Infusion Triptose (LIT) medium cultures. A simple, easy-to-execute and effective protocol based on ion exchange chromatography on Sepharose-DEAE resin for the purification of T. cruzi trypomastigotes is described. T. cruzi strains from the Discrete Typing Units (DTUs) I and II were used. The strains were harvested in LIT medium at a concentration of 1 × 107 epimastigotes/mL. We calculated the time of trypomastigotes increment (TTI). Based on the data obtained, Ion exchange chromatography was performed with DEAE-sepharose resin. To verify the purity and viability of the trypomastigotes, a culture was carried out in LIT medium with subsequent verification with giemsa staining. To evaluate if the technique affected the infectivity of trypomastigotes, in vitro assays were performed in Vero cells and in vivo in ICR-CD1 mice. The technique allowed the purification of metacyclic trypomastigotes of other stages of T. cruzi in a percentage of 100%, a greater recovery was observed in cultures of 12 days. There were differences regarding the recovery of metacyclic trypomastigotes for both DTUs, being DTU TcI the one that recovered a greater amount of these forms. The technique did not affect parasite infectivity in vitro or/and in vivo. © 2017 Elsevier B.V.eng
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.1016/j.mimet.2017.08.021
dc.identifier.issn1677012
dc.identifier.urihttps://repository.urosario.edu.co/handle/10336/22403
dc.language.isoengspa
dc.publisherElsevier B.V.spa
dc.relation.citationEndPage32
dc.relation.citationStartPage27
dc.relation.citationTitleJournal of Microbiological Methods
dc.relation.citationVolumeVol. 142
dc.relation.ispartofJournal of Microbiological Methods, ISSN:1677012, Vol.142,(2017); pp. 27-32spa
dc.relation.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85028709682&doi=10.1016%2fj.mimet.2017.08.021&partnerID=40&md5=a200932fa0d0a0952e55a5a66f5bf6fbspa
dc.rights.accesRightsinfo:eu-repo/semantics/openAccess
dc.rights.accesoAbierto (Texto Completo)spa
dc.source.instnameinstname:Universidad del Rosariospa
dc.source.reponamereponame:Repositorio Institucional EdocURspa
dc.subject.keywordSepharosespa
dc.subject.keywordDiethylaminoethyldextranspa
dc.subject.keywordinbred icreng
dc.subject.keywordion exchangeeng
dc.subject.keywordSepharosespa
dc.subject.keywordAnimal experimentspa
dc.subject.keywordArticlespa
dc.subject.keywordEpimastigotespa
dc.subject.keywordHost pathogen interactionspa
dc.subject.keywordIn vitro studyspa
dc.subject.keywordIn vivo studyspa
dc.subject.keywordIon exchange chromatographyspa
dc.subject.keywordLife cyclespa
dc.subject.keywordMousespa
dc.subject.keywordNonhumanspa
dc.subject.keywordParasitosisspa
dc.subject.keywordPriority journalspa
dc.subject.keywordTrypanosoma cruzispa
dc.subject.keywordTrypomastigotespa
dc.subject.keywordVero cell linespa
dc.subject.keywordAnimalspa
dc.subject.keywordCell linespa
dc.subject.keywordChlorocebus aethiopsspa
dc.subject.keywordHost pathogen interactionspa
dc.subject.keywordInstitute for cancer research mousespa
dc.subject.keywordIon exchange chromatographyspa
dc.subject.keywordIsolation and purificationspa
dc.subject.keywordProceduresspa
dc.subject.keywordTrypanosoma cruzispa
dc.subject.keywordAnimalsspa
dc.subject.keywordCell linespa
dc.subject.keywordCercopithecus aethiopsspa
dc.subject.keywordChromatographyeng
dc.subject.keywordDeae-dextranspa
dc.subject.keywordHost-pathogen interactionsspa
dc.subject.keywordMicespa
dc.subject.keywordMiceeng
dc.subject.keywordSepharosespa
dc.subject.keywordTrypanosoma cruzispa
dc.subject.keywordVero cellsspa
dc.subject.keywordChromatographyspa
dc.subject.keywordCulturespa
dc.subject.keywordMetacyclic trypomastigotesspa
dc.subject.keywordSepharose-deaespa
dc.subject.keywordTrypanosoma cruzispa
dc.titlePurification of Trypanosoma cruzi metacyclic trypomastigotes by ion exchange chromatography in sepharose-DEAE, a novel methodology for host-pathogen interaction studiesspa
dc.typearticleeng
dc.type.hasVersioninfo:eu-repo/semantics/publishedVersion
dc.type.spaArtículospa
Archivos
Bloque original
Mostrando1 - 1 de 1
Miniatura
Nombre:
1-s2-0-S0167701217302282-main.pdf
Tamaño:
890.6 KB
Formato:
Adobe Portable Document Format
Descripción:
Descargar
Colecciones
Artículos