Cultivation of Plasmodium falciparum Parasites in a Serum-Free Medium

Ofulla, Ayub V.; Okoye, Vincent; Khan, Baldip; Githure, John I.; Roberts, Clifford R.; Johnson, Anthony J.; Martin, Samuel K.

doi:10.4269/ajtmh.1993.49.335

Cited by 40 publications

(32 citation statements)

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“…Biochemical studies on blood-stage Plasmodium malarial parasites have demonstrated the parasite's proficiency at scavenging and modifying lipids obtained from the host [1]. Studies have also shown that the parasites could obtain free fatty acids (FFAs) directly from the serum or from sources such as high-density lipoprotein (HDL) [2–4]. Within the parasite, scavenged phospholipids can be incorporated without modification [3, 5].…”

Section: Introductionmentioning

confidence: 99%

Modulation of Lipoprotein Cholesterol Levels in Plasmodium berghei Malarial Infection by Crude Aqueous Extract of Ganoderma lucidum

et al. 2012

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In this study, attempt is made to establish changes in serum and liver lipoprotein cholesterols accompanying Plasmodium berghei malarial infection in mice treated with aqueous extract of Ganoderma lucidum at 100, 250, and 500 mg/kg body weight in comparison with 15 mg/kg chloroquine (CQ). Significant increases in all the lipoprotein fractions were observed in infected untreated mice compared with normal control mice. Treatment with 100 and 250 mg/kg G. lucidum extract produced significant reduction in serum total cholesterol (TC) and low-density cholesterol (LDL-C) contents compared with 500 mg/kg G. lucidum and CQ. Treatment with CQ, however, produced significant reduction in hepatic TC and LDL-C compared with the extract. A dose-dependent significant increase in serum high-density lipoprotein cholesterol (HDL-C) was observed in the G. lucidum treated mice compared with normal control but significantly lower compared with CQ-treated mice. Liver HDL-C level was significantly higher in CQ-treated mice compared with normal control and significantly lower compared with G. lucidum-treated and infected untreated mice. A dose-dependent effect of the extract was observed in both serum and liver very-low density lipoprotein cholesterol (VLDL-C). The implication of these results is discussed with respect to the parasite survival and proliferation in the serum and liver.

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Section: Introductionmentioning

confidence: 99%

Modulation of Lipoprotein Cholesterol Levels in Plasmodium berghei Malarial Infection by Crude Aqueous Extract of Ganoderma lucidum

et al. 2012

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“…Ifediba and Vanderberg (32) reported replacement of human serum by neopeptone and Proteose Peptone no. 3; bovine serum albumin (5 g/liter) was reported to replace serum by Ofulla et al (65). Dialyzed human serum lost its ability to support growth of parasites, and commercial samples of human serum supported growth at about one-quarter that of control cultures (37).…”

Section: Erythrocytic Stagesmentioning

confidence: 99%

Cultivation of Plasmodium spp

Schuster

2002

Clin Microbiol Rev

101

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Cultivation of both human and non-human species of Plasmodium spp., the causal agent of malaria, has been a major research success, leading to a greater understanding of the parasite. Efforts at cultivating the organisms in vitro are complicated by the parasites' alternating between a human host and an arthropod vector, each having its own set of physiological, metabolic, and nutritional parameters. Life cycle stages of the four species that infect humans have been established in vitro. Of these four, P. falciparum remains the only species for which all stages have been cultured in vitro; different degrees of success have been achieved with the other human Plasmodium spp. The life cycle includes the exoerythrocytic stage (within liver cells), the erythrocytic stage (within erythrocytes or precursor reticulocytes), and the sporogonic stage (within the vector). Culture media generally consist of a basic tissue culture medium (e.g., minimal essential medium or RPMI 1640) to which serum and erythrocytes are added. Most of the efforts have been directed toward the stage found in the erythrocyte. This stage has been cultivated in petri plates or other growth vessels in a candle jar to generate elevated CO2 levels or in a more controlled CO2 atmosphere. Later developments have employed continuous-flow systems to reduce the labor-intensive nature of medium changing. The exoerythrocytic and sporogonic life cycle stages have also been cultivated in vitro. A number of avian, rodent, and simian malarial parasites have also been established in vitro. Although cultivation is of great help in understanding the biology of Plasmodium, it does not lend itself to use for diagnostic purposes

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“…The continuous culture of P. falciparum in serum-free media is dependent upon the supply of exogenous purines (1,43), suggesting that the erythrocyte adenylate nucleotide pool is not a sufficient purine source and that the parasite is dependent upon extraerythrocytic purine salvage for survival. The first step in purine salvage is, therefore, the transport of purines into the infected cell and, from there, into the intracellular parasite.…”

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Purine Salvage Pathways in the Intraerythrocytic Malaria Parasite Plasmodium falciparum

2008

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2Malaria is caused by protozoan parasites of the genus Plasmodium. Five species of Plasmodium cause infection in humans, with the majority of lethal cases caused by Plasmodium falciparum. This species is responsible for more than 500 million clinical cases of malaria each year (59). In the past 2 decades, efforts to combat this disease have been met with the emergence of widespread resistance to most of the commonly used antimalarial drugs (68). The poor efficacy of current drugs and the lack of a promising vaccine have resulted in alarming increases in the rates of malaria morbidity and mortality worldwide, with the major toll felt in the developing world.P. falciparum is transmitted to humans via the bite of an infected female anopheline mosquito. In humans, the parasite undergoes one cycle of asexual multiplication in hepatocytes, followed by several cycles of infection and multiplication in red blood cells. Whereas the hepatocytic stage is asymptomatic, the erythrocytic stage is accompanied by the destruction of the host erythrocytes, resulting in anemia and, in the absence of treatment, death. Extensive efforts are presently under way to develop a vaccine, and advances in our understanding of the biology of the parasite and its metabolic and nutritional needs offer new routes for chemotherapy. In this regard, purine metabolism holds significant promise as a target for drug development.It has long been recognized that protozoan parasites, including Plasmodium spp., are unable to synthesize purine rings de novo (4). Consistent with this observation, the sequencing of protozoan genomes has failed to uncover any genes encoding enzymes involved in the biosynthesis of purine nucleosides or nucleobases (12). Protozoan parasites rely instead on salvage of purines from the host. The strategies used in acquiring purines vary significantly among parasite genera. This review will focus primarily on the purine salvage pathways present in the Plasmodium parasite. Recent reviews have examined the metabolic pathways for purine salvage in other protozoa (11,17,29).Initial studies on purine and pyrimidine synthesis in Plasmodium parasites were performed on erythrocytic stages of the rodent malaria species P. berghei (7,8,65), the macaque monkey malaria species P. knowlesi (46), and the avian malaria parasite P. lophurae (61,66 (46). Following on from these findings, Gutteridge and Trigg found that, in P. knowlesi, all radiolabeled purines were significantly incorporated into nucleic acids while none of the pyrimidines tested were (28). These early biochemical studies demonstrated that Plasmodium parasites lack the ability to metabolize exogenous pyrimidines and instead are entirely dependent on de novo synthesis. Conversely, Plasmodium parasites are entirely reliant upon the salvage of extracellular purines (4) and are capable of metabolizing a wide variety of exogenous purine nucleobases and nucleosides.The continuous culture of P. falciparum in serum-free media is dependent upon the supply of exogenous purines (1, 43), sug...

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Cultivation of Plasmodium falciparum Parasites in a Serum-Free Medium

Cited by 40 publications

References 0 publications

Modulation of Lipoprotein Cholesterol Levels in Plasmodium berghei Malarial Infection by Crude Aqueous Extract of Ganoderma lucidum

Modulation of Lipoprotein Cholesterol Levels in Plasmodium berghei Malarial Infection by Crude Aqueous Extract of Ganoderma lucidum

Cultivation of Plasmodium spp

Purine Salvage Pathways in the Intraerythrocytic Malaria Parasite Plasmodium falciparum

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