Efficiency of in vitro culture of Toxoplasma gondii in THP1 and Vero cell lines
Abstract
Introduction. Cell culture is an important method for isolating Toxoplasma gondii to make clinicaldiagnosis or for biotechnological purposes.
Objective. The percentage of invasion and production levels of T. gondii was determined for THP1 andVero cell lines.
Materials and methods. The growth conditions for T. gondii in Vero and THP1 cell lines were determinedby counting in hemocytometer chamber. The percentage of invasion of T. gondii in THP1 and Verocells was determined by flow cytometry in different cell/tachyzoite ratios: 1/5, 1/20, 1/50. The growthperformance index of the T. gondii RH strain and the CIBM1 isolate was calculated for THP1 cells.
Results. Vero cells multiplied faster than the THP1 cells, showing an exponential and a sigmoidalgrowth curve respectively, within a period of 7 days. The CIBM1 isolate infected the THP1 cells in threedifferent parasite concentrations: 1/5, 1/20 and 1/50, with invasion percentages in THP1 cells of 57.1%,15.5% and 12.2% and for the Vero cells 25.3%, 17.8% and 8.8% respectively. The RH strain of T. gondiihad the lowest invasion percentage with 32.6%, 14.8% and 8.1% in THP1 cells and 22.3%, 14.1% and3.4% in Vero cells.
Conclusions. The CIBM1 isolate had a higher yield than the RH strain of T. gondii in THP1 cells.THP1 cells were indicated to be a good model for the study of invasion and for the assays of newpharmacological candidates against T. gondii.
doi: http://dx.doi.org/10.7705/biomedica.v32i3.485
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References
Adl SM, Simpson AG, Farmer MA, Andersen RA, Anderson OR, Barta JR, et al. The new higher level classification of eukaryotes with emphasis on the taxonomy of protists. J Eukaryot Microbiol. 2005;5:399-451. http://dx.doi.org/10.1111/j.1550-7408.2005.00053.x
Smith JR, Franc DT, Carter NS, Zamora D, Planck SR, Rosenbaum JT. Susceptibility of retinal vascular endothelium to infection with Toxoplasma gondii tachyzoites. Invest Ophthalmol Vis Sci. 2004;45:1157-61. http://dx.doi.org/10.1167/iovs.03-1105
Gil A, Ramírez MC, Aguilera MC, Mesa MD. Modelos experimentales de enfermedad cardiovascular. Nutr Hosp. 2007;22:169-77.
Gizem S, Emin ÜB. Production of Toxoplasma gondii in Vero Cell Culture. Turkiye Parazitol Derg. 2011;35:61-4.
Shapiro HM. Practical flow cytometry. Fourth edition. Hoboken, NJ: John Wiley & Sons, Inc.; 2003.
Bouix M, Leveau JY. The applications of flow cytometry in microbiology. Bull Soc Fr Microbiol. 2001;16:210-8.
Bouchot A, Millot MJ, Charpentier S, Bonhomme A, Villena I, Aubert D, et al. Membrane potential changes after infection of monocytes by Toxoplasma gondii. Int J Parasitol. 2001;31:1114-20. http://dx.doi.org/10.1016/S0020-7519(01)00224-7
Saadatnia G, Haj Ghani H, Khoo BY, Maimunah A, Rahmah N. Optimization of Toxoplasma gondii cultivation in Vero cell line. Trop Biomed. 2010;27:125-30.
De Meerschman F, Rettigner C, Focant C, Boreux R, Pinset C, Leclipteux T, et al. Use of a serum-free medium to produce in vitro Neospora caninum and Toxoplasma gondii tachyzoites on Vero cells. Vet Res. 2002;33:159-68.http://dx.doi.org/10.1051/vetres:2002004
Degirmenci A, Doskaya M, Caner A, Cicek C, Korkmaz M, Guruz Y, et al. Toxoplasma gondii RH Ankara: Production of evolving tachyzoites using a novel cell culture method.
Exp Parasitol. 2011;128:1-8. http://dx.doi.org/10.1016/j.exppara.2011.01.019
Appleford JP, Smith EJ. Toxoplasma gondii: The growth characteristics of three virulent strains. Acta Tropica. 1997;65:97-104.http://dx.doi.org/10.1016/S0001-706X
(97)00656-6
Ashburn D, Evans R, Chatterton JM, Joss AW, Ho-Yen DO. Toxoplasma dye test using cell culture derived tachyzoites. J Clin Pathol. 2003;53:630-3. http://dx.doi.org/10.1136/jcp.53.8.630
Chatterton JM, Evans R, Ashburn D, Joss AW, Ho-Yen DO. Toxoplasma gondii in vitro culture for experimentation.J Microbiol Methods. 2002;51:331-5. http://dx.doi.
org/10.1016/S0167-7012(02)00101-X
Evans R, Chatterton JM, Ashburn D, Joss AW, Ho-Yen DO. Cell-culture system for continuous production of Toxoplasma gondii tachyzoites. Eur J Clin Microbiol.1999;18:879-84.
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