Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 32578
Plaque Formation of Toxoplasma gondii in Vero Cells using Carboxymethylcellulose

Authors: L. Fonseca-Géigel, M. Alvarez, G. García, R. Cox, L. Morier, L. Fonte, M. G. Guzmán


Toxoplasma gondii is an intracellular parasite capable of infecting all nucleated cells in a diverse array of species. Toxoplasma plaque assay have been described using Bacto Agar. Because of its experimental advantages carboxymethyl cellulose overlay, medium viscosity was choosing and the aim of this work was to develop alternative method for formation of T. gondii plaques. Tachyzoites were inoculated onto monolayers of Vero cells and cultured at 37° C under 5 % CO2. The cultures were followed up by microscopy inspection. Small plaques were visible by naphtol blue stain 4 days after infection. Larger plaques could be observed by day 10 of culture. The carboxymethyl cellulose is a cheap reagent and the methodology is easier, faster than assays under agar overlay. This is the first description of the carboxymethyl cellulose overlay use for obtaining the formation of T. gondii plaques and may be useful in consequent obtaining tachyzoites for detailed studies.

Keywords: Carboxymethyl cellulose, Cell culture, Plaque assay, Toxoplasma gondii.

Digital Object Identifier (DOI):

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2647


[1] J.G. Montoya and O. Liesenfeld, "Toxoplasmosis", 2004, The Lancet 363, pp. 1965-1976.
[2] Chatterton, R. Evans, D. Ashburn, A.W.L. Joss and D.O. Ho-Yen, "Toxoplasma gondii in vitro culture for experimentation", 2002, Journal of Microbiology Methods 51, pp. 331-335.
[3] X. Li, Y. Hao, D. Chen, Q. Liu, J. Ding and W. Zhang, "Expression of the tandem enhanced yellow fluorescent marker gene in Toxoplasma gondii", 2009, Parasitology Research 105, pp. 287-291.
[4] D.S. Roos, R.G. Donald, N.S. Morrissette and A.L. Moulton, "Molecular tools for genetic dissection of the protozoan parasite Toxoplasma gondii", 1994, Methods in Cell Biology 45: pp. 27-63.
[5] E.R. Pfefferkorn and L.C. Pfefferkorn, "Toxoplasma gondii: isolation and preliminary characterization of temperature sensitive mutants", 1976, Experimental Parasitology 39: pp. 365-376.
[6] P.K. Russell, S. Udomsakdi and S.B. Halstead, "Antibody response in dengue and dengue hemorrhagic fever", 1967, Japan Journal of Medicine Science Biology 20: Supp:103-8.
[7] M. Matrosovich, T. Matrosovich, W. Garten and H.D. Klenk, "New lowviscosity overlay medium for viral plaque assays", 2006, Virology Journal 3: pp. 63. Epub Aug 31, 2006. DOI: 10.1186/1743-422X-3-63.
[8] P. Herzog, C. Drosten and M. Muller, "Plaque assay for human coronavirus NL63 using human colon carcinoma cells", 2008, Virology Journal 5: pp. 138. Epub Nov 12, 2008. DOI: 10.1186/1743-422X-5- 138.
[9] S.D. Chaparas and R.W. Schlesinger, "Plaque assay of Toxoplasma on monolayer of chick embryo fibroblasts", 1959, Proceedings of the Society for Experimental Biology and Medicine 102: pp. 431-437.
[10] D.H. Hollander, "Colonial morphology of Trichomonas vaginalis in agar", 1976, Journal of Parasitology 62: pp. 826-828.
[11] D.E. Mueller and W.A. Petri, "Clonal growth in petri dishes of Entamoeba histolytica", 1995, Transaction of the Royal Society of Tropical Medicine and Hygiene 89: pp. 123.
[12] E.M. Valido and W.L. Rivera, "Colony growth of philippine isolates of Blastocystis hominis in simplified, soft agar medium", 2007, Parasitology Research 101: pp. 213-217.
[13] M. Hess, T. Kolbe, E. Grabensteiner and H. Prosl, "Clonal cultures of Histomonas meleagridis, Tetratrichomonas gallinarum and a Blastocystis sp. established through micromanipulation", 2006, Parasitology 133: pp. 547-554.
[14] S.A. Edgar and C.T. Seibold, "A new coccidium of chickens, Eimeria mivati sp. n. (Protozoa: Eimeriidae) with details of its life history", 1964, Journal of Parasitology 500: pp. 193-204.
[15] J.R. Sreekumar, A.K. Rao, A. D. Mishra, G.C. Ray and G.C. Bansal, "In vitro generation of cloned populations of Toxoplasma gondii", 2003, Parasitology Research 90: pp. 489-492.