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Scientific and technological research article

Vol. 15 No. 2 (2013)

Comparison of "in vitro" culture of Toxoplasma gondii RH strain in Hep-2 and Vero cell lines

Submitted
November 11, 2015
Published
2013-11-05

Abstract

Introduction: Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, which is an obligate intracellular parasite capable of replicate itself in all nucleated cells, allowing their isolation and maintenance in cell culture. Objective: To compare cultivation Toxoplasma gondii (RH) in Hep-2 cells and Vero employed to spread parasitic. Methods: The conditions were optimized for cultivation of Toxoplasma gondii (RH) in Vero and Hep-2 cells lines, 500.000 and 1.000.000cel./mm3 were seeded and 1.000.000 and 2.000.000taq/mm3 were infected respectively, a track to determine cell invasion, percentage of rosettes and viability and efficiency of tachyzoites was made until the 120 hours. Results: After 48 hours of incubation, 90% of parasite invasion was evident in both cell lines as well as the largest count rosettes. At 120 hours higher counts of extracellular tachyzoites and a higher rate of performance of 185.9 in Hep-2 cells at concentration of 1.000.000taq/mm3 and 500.000cel./mm3 were obtained. Average parasite viability over 80 % was obtained for both cell lines. Conclusions: Higher parasitic performance was obtained viable in Hep-2 cells, so that this cell line can be considered as the preferred model for the propagation and maintenance of T. gondii (RH).

References

1. Peña Y, Bernal R, Pavía N. Toxoplasmosis diseminada en niños. informe de dos casos). Enf Infec y Micro. 2003;23:149-54.

2. Lindsay D, Blackburn B, Dubey J. Feline toxoplasmosis and the importance of the Toxoplasma gondii oocyst. Parasitol. 1997;19:448-61.

3. Wong S, Remington J. Biology of Toxoplasma gondii. AIDS. 1993; 7:299-316.

4. Cuellar J, Hernández A, Villegas E, Gómez J. Eficiencia de cultivo in vitro de Toxoplasma gondii en las líneas celulares THP1 y Vero. Biomédica.
2012;32:461-66 doi: http://dx.doi.org/10.7705/biomedica.v32i3.485.

5. Gomes T, de Andrade Júnior H, Zevallos S, Amato-Neto V. In vitro action of antiparasitic drugs, especially artesunate, against Toxoplasma gondii. Rev Soc Bras Med Trop. 2012;45:485-90.

6. James G, Sintchenko V, Dickeson D, Gilbert Gl. Comparison of Cell Culture, Mouse Inoculation, and PCR for Detection of Toxoplasma gondii: Effects of Storage Conditions on Sensitivity. J. Clin. Microbiol. 1996;34:1572–75

7. Cooper G, Hausman R, Wrigth N. La célula. 5 ed. Madrid: Marbán Libros; 2010.

8. Freshney R. Culture of animal cells. A manual of Basic Technique. 3 ed. Glassgow: Editorial John Wiley & Sons; 2005.

9. Harmer C, Hassl A, Kreinecker S, Aspöck H. Toxoplasma gondii in vitro cultivation: economic and efficient mass production. J. Microbiol. Methods.1996;27:225-28.

10. Harmer C, Aspöck H, Hassl A. Toxoplasma gondii in vitro cultivation: easy handling long-term propagation. J. Microbiol. Methods. 1996; 27:221-23.

11. Chatterton J, Evans R, Ashburn D, Joss A, Ho-Yen D. Toxoplasma gondii in vitro culture for experimentation. J. Microbiol. Methods. 2002;51:331-35.

12. Osorio A. Ética en la investigación con modelos animales experimentales. Alternativas y las 3 RS de Russel. Una responsabilidad y un compromiso ético que nos compete a todos. Rev. Col. Bioética. 2006; 163-83.

13. Degirmenci A, Doskaya M. Ayse C, Candan Ç, Metin K, Yüksel G, et al. Toxoplasma gondii RH Ankara: Production of evolving tachyzoites using a novel cell culture method. Exp. Parasitol. 2011;128:1-8.

14. 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.

15. Gallego C, Castaño JC, Giraldo A, Ajzenberg D, Dardé ML, Gómez JE. Caracterización biológica y molecular del aislado CIBMUQ/HDC una cepa Colombiana de referencia para Toxoplasma gondii. Biomédica. 2004;24:282-90.

16. Jones-Brando L, D’Angelo J, Posner G, Yolken R. In vitro inhibition of Toxoplasma gondii by four new derivatives of artemisinin. Antimicrob Agents Chemother. 2006; 50: 4206–08. doi: 10.1128/AAC.00793-06.

17. Sivrikaya G, Bağrıaçık EÜ. Production of Toxoplasma gondii in Vero cell culture. Turkiye Parazitol Derg. 2011;35:61-4. doi: 10.5152/tpd.2011.16.

18. Canessa M, Melioli G. Enzyme Immunoassay for Evaluation of Toxoplasma gondii growth in Tissue Culture. J. Clin. Microbiol. 1984;21:88-91.

19. Russell W, Burch R. The principles of Humane Experimental Technique, London; Methuen & Co Ltda; 1959.

20. Ding J, Wu K, Tan F, Chen XG. Establishment of an in vitro tachyzoite-bradyzoite interconversion system for Toxoplasma gondii. Nan Fang Yi Ke Da Xue Xue Bao. 2010;30:668-71.

21. Evans R, Chatterton J, Ashburn D, Joss A, Ho-Yen DO. Cell-culture system for continuous production of Toxoplasma gondii tachyzoites. Eur J Clin Microbiol Infect Dis. 1999;18:879-84.
22. Diab M. El-Bahy M. Toxoplasma gondii: Virulence of tachyzoites in serum free media at different temperatures. Exp. Parasitol. 2008;118:75-79.

23. Bilgin M, Yıldırım T, Hökelek Balkan M. In Vitro Effects of Ivermectin and Sulphadiazine on Toxoplasma gondii. Med. J. 2013;30:19-22 doi: 10.5152/balkanmedj. 2012.098.

24. Dauci H, Ciler A, Uner A, Aksoy U, Ustun S. In vivo and in vitro effect of Clarithromycin on Toxoplasma gondii. Turk Journal Inf. 2002;17:325-28.

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