Toxoplasma gondii RH Ankara: Production of evolving tachyzoites using a novel cell culture method

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dc.contributor.authorDegirmenci, Aysu
dc.contributor.authorDoskaya, Mert
dc.contributor.authorCaner, Ayse
dc.contributor.authorCicek, Candan
dc.contributor.authorKorkmaz, Metin
dc.contributor.authorGuruz, Yuksel
dc.contributor.authorUner, Ahmet
dc.date.accessioned2019-10-27T21:24:24Z
dc.date.available2019-10-27T21:24:24Z
dc.date.issued2011
dc.departmentEge Üniversitesien_US
dc.description.abstractToxoplasma gondii is one of the most researched parasite due to its easy growth both in vitro and in vivo. Tachyzoites, derived from mouse or rat peritoneum encounters ethical and economical problems when used for research or diagnostic purposes. Currently, research has focused on determining the most suitable cell culture environment to reach highest amount of viable tachyzoites with least host cell contamination. However, gene expression changes that take place throughout the adaptation of evolving T. gondii strains to continuous cell cultures appear as a problem. The present study aimed to determine a novel cell culture strategy for T. gondii RH Ankara strain tachyzoites to harvest abundant tachyzoites with least host cell contamination and minimal antigenic variation at predetermined dates to use as an antigen source in serological assays that will facilitate reduction in animal use. To achieve this purpose, T. gondii RH Ankara strain tachyzoites were incubated with HeLa cell at different ratios for two or three days. In all flasks incubated for two days, viability rate reached to 100% and HeLa cell contamination decreased to levels between 0.12-0.5 x 10(6)/ml. In the flasks with HeLa-tachyzoite ratio 1/8, the tachyzoite yield and viability ratio were 3 x 10(6)/ml and 100%, respectively, with accompanying 10 fold decrease (0.12 x 10(6)/ml) in HeLa contamination. During continuous production, highest tachyzoite yield was obtained from the first passage (3.55 x 10(6)/ml) and until the end of third subculture viability rates and HeLa cell contaminations were between 98.2-99.4% and 0.31-0.37 x 10(6)/ml, respectively. ELISA. IFA and Western blot analyses showed that the quality, specificity and sensitivity of the antigen harvested from the first passage of cell culture performed at two days intervals were comparable to the antigen harvested from mice and decreased in the following subcultures. Overall, these results demonstrated that T. gondii RH Ankara strain is still evolving to adapt to cell culture environment and therefore such strains continuously produced in cell cultures should be avoided for serological assays. However, the two day short interval cell culture method described herein offers a chance to reduce the animal use intended for the preparation of serological assays' antigen from local evolving strains. (C) 2011 Elsevier Inc. All rights reserved.en_US
dc.description.sponsorshipScientific and Technical Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [SBAG 105S022]; Ege University Science and Technology Centre (EBILTEM)Ege University [2006 BIL 012]en_US
dc.description.sponsorshipThe research was supported by the grant given by Scientific and Technical Research Council of Turkey (TUBITAK) (Project no: SBAG 105S022) and partially by the grant given by Ege University Science and Technology Centre (EBILTEM) (Project no: 2006 BIL 012).en_US
dc.identifier.doi10.1016/j.exppara.2011.01.019en_US
dc.identifier.endpage8en_US
dc.identifier.issn0014-4894
dc.identifier.issn1090-2449
dc.identifier.issue1en_US
dc.identifier.pmid21296077en_US
dc.identifier.scopusqualityQ2en_US
dc.identifier.startpage1en_US
dc.identifier.urihttps://doi.org/10.1016/j.exppara.2011.01.019
dc.identifier.urihttps://hdl.handle.net/11454/44523
dc.identifier.volume128en_US
dc.identifier.wosWOS:000289034700001en_US
dc.identifier.wosqualityQ3en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherAcademic Press Inc Elsevier Scienceen_US
dc.relation.ispartofExperimental Parasitologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectEvolving Toxoplasma gondii strainsen_US
dc.subjectCell cultureen_US
dc.subjectHeLaen_US
dc.subjectAntigenen_US
dc.titleToxoplasma gondii RH Ankara: Production of evolving tachyzoites using a novel cell culture methoden_US
dc.typeArticleen_US

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