Successful in vitro culture of Plasmodium falciparum gametocytes
Keywords:
Plasmodium falciparum, in vitro, malaria, Anopheles
Abstract
Introduction. The sexual stages of Plasmodium falciparum have not been studied in as much detail as the asexual stages due to the lack of standardized in vitro cultures as well as difficulties in identifying the sexual development stages of the parasite. These difficulties hamper the studies on biology, metabolism, gene expression and protein synthesis during sexual stages. Each of these facets are important targets in antimalarial drug research, particularly the identification of potential therapeutic agents against Plasmodium (derived mainly from plants).Objectives. An in vitro culture of P. falciparum gametocytes was established to standardize the identification of its five developmental stages and ensure their continuous production.
Materials and methods. The in vitro gametocyte culture was established from the P. falciparum NF54 strain in RPMI culture medium, with assessment of the asexual and sexual parasitaemia. The medium was supplemented with type A Rh+ red blood cells only on the first day of culture. Subsequently, the medium was changed daily, together with addition of gas mixture (90% N2, 5% O2, 5% CO2) and maintenance of the culture temperature at 37 °C. When asexual parasitaemia reached 3 to 5%, the medium was changed by doubling its volume.
Conclusions. We standardized an in vitro culture for sexual stages of P. falciparum that can be used for future studies about evaluation of compounds of synthetic or natural origen against the sexual stage, which may permit to develop new control strategies against malaria.
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References
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4. Sinden RE. Sexual development of malaria parasite. Adv Parasitol. 1983;22:153-216.
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9. Hawking F, Wilson ME, Gammage K. Evidence for cyclic short-lived maturity in the gametocytes of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 1971;65:549-59.
10. Field JW, Shute PG. The microscopic diagnosis of human malaria. A morphological study of erythrocytic parasites. Kuala Lumpur, Malaysia: Institute for Medical Research, Government Press; 1956. p. 142.
11. Lobo CA, Kumar N. Sexual differentiation and development in the malaria parasite. Parasitol Today. 1998;14:146-50.
12. Row R. On some observation on the malarial parasites grow aerobically in simple cultures with special reference to the evolution and degeneration of the crescents. Indian J Med Res. 1929;16:120-7.
13. Trager W, Jensen JB. Human malaria parasite in continuous culture. Science. 1976;193:673-5.
14. Smalley ME. Plasmodium falciparum gametocytogenesis in vitro. Nature. 1976;264:271-2.
15. Ifediba T, Vanderberg JP. Complete in vitro maturation of Plasmodium falciparum gametocytes. Nature. 1981;294:364-6.
16. Ponnudurai T, Meuwissen JH, Leeuwenberg AD, Verhave JP, Lensen AH. The production of mature gametocytes of Plasmodium falciparum in continuous cultures of different isolates infective to mosquitoes. Trans R Soc Trop Med Hyg. 1982;76:242-50.
17. Freese JA, Sharp BL, Ridl FC, Markus MB. In vitro cultivation of southern African strains of Plasmodium falciparum and gametocytogenesis. S Afr Med J .1988;73:720-2.
18. Ponnudurai T, Lensen AH, Meuwissen JH. An automated large – scale culture system of Plasmodium falciparum using tangential flow filtration for medium change. Parasitology. 1983;87:439-45.
19. Ponnudurai T, Lensen AH, Meis JF, Meuwissen JH. Synchronization of Plasmodium falciparum gametocytes using an automated suspension culture system. Parasitology. 1986;93:263-74.
20. Kanti BM, Kumar N. Plasmodium falciparum gametocyte culture, purification, and gametogenesis. In: Ljungström I, Perlmann H, Schlichtherle M, Scherf A, Wahlgren M, editors. Methods in malaria research. Fourth edition. Manassas, Virginia; MR4/ATCC; 2004. p. 93-4.
21. Instituto Nacional de Salud. Protocolos para el manejo de la cepa de P. falciparum in vitro. Bogotá. Instituto Nacional de Salud; 1989. p. 28.
22. Ali E, Mackinnon MJ, Abdel-Muhsin AM, Ahmed S, Walliker D, Babiker HA. Increased density but not prevalence of gametocytes following drug treatment of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 2006;100:176-83.
23. Sutherland CJ, Alloueche A, Curtis J, Drakeley CJ, Ord R, Duraisingh M, et al. Gambian children successfully treated with chloroquine can harbor and transmit Plasmodium falciparum gametocytes carrying resistance genes. Am J Trop Med Hyg. 2000;67:578-85.
24. Sinden RE, Ponnudurai T, Smits MA, Simm AM, Meuwissen JH. Gametocytogenesis of Plasmodium falciparum in vitro: a simple technique for the routine culture of pure capacitated gametocytes en masse. Parasitology. 1984;88:239-47.
25. Chutmongkonkul M, Maier WA, Seitz HM. A new model for testing gametocytocidal effects of some antimalarial drugs on Plasmodium falciparum in vitro. Ann Trop Med Parasitol. 1992;86:207-15.
26. Saul A, Graves P, Edser L. Refractoriness of erythrocytes infected with Plasmodium falciparum gametocytes to lysis by sorbitol. Int J Parasitol. 1990;20:1095-7.
27. Kariuki MM, Kiaira JK, Mulaa FK, Mwangi JK, Wasunna MK, Martin SK. Plasmodium falciparum: Purification of the various gametocyte developmental stages from in vitro–cultivated parasites. Am J Trop Med Hyg. 1998;59:505-8.
28. Jensen JB. Observations on gametogenesis in Plasmodium falciparum from continuous culture. J Protozool. 1979;26:129-32.
29. Phillips RS, Wilson RJ, Pasvol G. Differentiation of gametocytes in microcultures of human blood infected with Plasmodium falciparum. J Protozool. 1978;25:394-8.
2. Shute PG, Maryon M. A study of gametocytes in a West African strain of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 1951;44:421-38.
3. Eichner M, Diebner HH, Molineaux L, Collins WE, Jeffery GM, Dietz K. Genesis, sequestration and survival of Plasmodium falciparum gametocytes: parameter estimates from fitting a model to malaria therapy data. Trans R Soc Trop Med Hyg. 2001;95:497-501.
4. Sinden RE. Sexual development of malaria parasite. Adv Parasitol. 1983;22:153-216.
5. Kitchen SF, Putnam P. Observations on the mechanism of the parasite cycle in falciparum malaria. Am J Trop Med. 1942; 22:361-86.
6. Kitchen SF. Falciparum malaria. En: Boyd MF, editor. Malariology. Philadelphia: WB Saunders; 1949. p. 995- 1016.
7. Carter R, Graves PM. Gametocytes. En: Wernsdorfer WH, McGregor SI, editors. Malaria: principles and practice of malariology. First edition. Edinburgh, Scotland: Churchill Livingstone; 1988. p. 1-59.
8. Talman AM, Domarle O, Mckenzie FE, Ariey F, Robert V. Gametocytogenesis: the puberty of Plasmodium falciparum. Malar J. 2004;3:24.
9. Hawking F, Wilson ME, Gammage K. Evidence for cyclic short-lived maturity in the gametocytes of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 1971;65:549-59.
10. Field JW, Shute PG. The microscopic diagnosis of human malaria. A morphological study of erythrocytic parasites. Kuala Lumpur, Malaysia: Institute for Medical Research, Government Press; 1956. p. 142.
11. Lobo CA, Kumar N. Sexual differentiation and development in the malaria parasite. Parasitol Today. 1998;14:146-50.
12. Row R. On some observation on the malarial parasites grow aerobically in simple cultures with special reference to the evolution and degeneration of the crescents. Indian J Med Res. 1929;16:120-7.
13. Trager W, Jensen JB. Human malaria parasite in continuous culture. Science. 1976;193:673-5.
14. Smalley ME. Plasmodium falciparum gametocytogenesis in vitro. Nature. 1976;264:271-2.
15. Ifediba T, Vanderberg JP. Complete in vitro maturation of Plasmodium falciparum gametocytes. Nature. 1981;294:364-6.
16. Ponnudurai T, Meuwissen JH, Leeuwenberg AD, Verhave JP, Lensen AH. The production of mature gametocytes of Plasmodium falciparum in continuous cultures of different isolates infective to mosquitoes. Trans R Soc Trop Med Hyg. 1982;76:242-50.
17. Freese JA, Sharp BL, Ridl FC, Markus MB. In vitro cultivation of southern African strains of Plasmodium falciparum and gametocytogenesis. S Afr Med J .1988;73:720-2.
18. Ponnudurai T, Lensen AH, Meuwissen JH. An automated large – scale culture system of Plasmodium falciparum using tangential flow filtration for medium change. Parasitology. 1983;87:439-45.
19. Ponnudurai T, Lensen AH, Meis JF, Meuwissen JH. Synchronization of Plasmodium falciparum gametocytes using an automated suspension culture system. Parasitology. 1986;93:263-74.
20. Kanti BM, Kumar N. Plasmodium falciparum gametocyte culture, purification, and gametogenesis. In: Ljungström I, Perlmann H, Schlichtherle M, Scherf A, Wahlgren M, editors. Methods in malaria research. Fourth edition. Manassas, Virginia; MR4/ATCC; 2004. p. 93-4.
21. Instituto Nacional de Salud. Protocolos para el manejo de la cepa de P. falciparum in vitro. Bogotá. Instituto Nacional de Salud; 1989. p. 28.
22. Ali E, Mackinnon MJ, Abdel-Muhsin AM, Ahmed S, Walliker D, Babiker HA. Increased density but not prevalence of gametocytes following drug treatment of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 2006;100:176-83.
23. Sutherland CJ, Alloueche A, Curtis J, Drakeley CJ, Ord R, Duraisingh M, et al. Gambian children successfully treated with chloroquine can harbor and transmit Plasmodium falciparum gametocytes carrying resistance genes. Am J Trop Med Hyg. 2000;67:578-85.
24. Sinden RE, Ponnudurai T, Smits MA, Simm AM, Meuwissen JH. Gametocytogenesis of Plasmodium falciparum in vitro: a simple technique for the routine culture of pure capacitated gametocytes en masse. Parasitology. 1984;88:239-47.
25. Chutmongkonkul M, Maier WA, Seitz HM. A new model for testing gametocytocidal effects of some antimalarial drugs on Plasmodium falciparum in vitro. Ann Trop Med Parasitol. 1992;86:207-15.
26. Saul A, Graves P, Edser L. Refractoriness of erythrocytes infected with Plasmodium falciparum gametocytes to lysis by sorbitol. Int J Parasitol. 1990;20:1095-7.
27. Kariuki MM, Kiaira JK, Mulaa FK, Mwangi JK, Wasunna MK, Martin SK. Plasmodium falciparum: Purification of the various gametocyte developmental stages from in vitro–cultivated parasites. Am J Trop Med Hyg. 1998;59:505-8.
28. Jensen JB. Observations on gametogenesis in Plasmodium falciparum from continuous culture. J Protozool. 1979;26:129-32.
29. Phillips RS, Wilson RJ, Pasvol G. Differentiation of gametocytes in microcultures of human blood infected with Plasmodium falciparum. J Protozool. 1978;25:394-8.
How to Cite
1.
Blair S, Rada AM, Moreno C. Successful in vitro culture of Plasmodium falciparum gametocytes. Biomed. [Internet]. 2008 Dec. 1 [cited 2025 Apr. 7];28(4):607-15. Available from: https://revistabiomedicaorg.biteca.online/index.php/biomedica/article/view/67
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