An alternative to serum for cultivation of Plasmodium falciparum in vitro

Cranmer, Susan L.; Magowan, Cathleen; Liang, Jie; Coppel, Ross L.; Cooke, Brian M.

doi:10.1016/s0035-9203(97)90110-3

Cited by 193 publications

(154 citation statements)

References 15 publications

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“…Erythrocytic stages of P. falciparum 3D7 isolate were cultured as described (33,34). The culture was checked monthly for Mycoplasma contamination (PCR Mycoplasma Test kit; MDB, St. Paul, MN).…”

Section: Methodsmentioning

confidence: 99%

Malaria hemozoin is immunologically inert but radically enhances innate responses by presenting malaria DNA to Toll-like receptor 9

Parroche

Lauw

Goutagny

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

482

459

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Hemozoin (HZ) is an insoluble crystal formed in the food vacuole of malaria parasites. HZ has been reported to induce inflammation by directly engaging Toll-like receptor (TLR) 9, an endosomal receptor. ''Synthetic'' HZ (␤-hematin), typically generated from partially purified extracts of bovine hemin, is structurally identical to natural HZ. When HPLC-purified hemin was used to synthesize the crystal, ␤-hematin had no inflammatory activity. In contrast, natural HZ from Plasmodium falciparum cultures was a potent TLR9 inducer. Natural HZ bound recombinant TLR9 ectodomain, but not TLR2. Both TLR9 stimulation and TLR9 binding of HZ were abolished by nuclease treatment. PCR analysis demonstrated that natural HZ is coated with malarial but not human DNA. Purified malarial DNA activated TLR9 but only when DNA was targeted directly to the endosome with a transfection reagent. Stimulatory quantities of natural HZ contain <1 g of malarial DNA; its potency in activating immune responses was even greater than transfecting malarial DNA. Thus, although the malarial genome is extremely AT-rich, its DNA is highly proinflammatory, with the potential to induce cytokinemia and fever during disease. However, its activity depends on being bound to HZ, which we propose amplifies the biological responses to malaria DNA by targeting it to a TLR9 ؉ intracellular compartment.fever ͉ immunomodulator ͉ parasitic diseases

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Section: Methodsmentioning

confidence: 99%

Malaria hemozoin is immunologically inert but radically enhances innate responses by presenting malaria DNA to Toll-like receptor 9

Parroche

Lauw

Goutagny

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

482

459

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“…However, the development of P. falciparum requires the presence of currently unknown factors present in human serum (Jensen, 1979). Although numerous studies have attempted to identify the factors and substances able to sustain parasite growth (Asahi & Kanazawa, 1994;Asahi et al, 1996Asahi et al, , 2005Cranmer et al, 1997;Divo and Jensen. 1982;Lingnau et al, 1994;Mi-Ichi et al, 2006;Nivet et al, 1983;Ofulla et al, 1993;Willet and Canfield, 1984), the establishment of a fully-defined culture medium for the parasite has represented a major challenge.…”

Section: Cdm For Continuous Intraerythrocytic Growth Of P Falciparummentioning

confidence: 99%

“…GFS contains lipid-rich albumin as a major component. Similarly, Cranmer et al (1997) described a commercially available lipidenriched bovine albumin (Albumax II; Invitrogen) that could replace human serum for the in vitro cultivation of P. falciparum. These serum substitutes are currently widely employed to maintain parasite cultures.…”

Section: Cdm For Continuous Intraerythrocytic Growth Of P Falciparummentioning

confidence: 99%

“…The mechanisms responsible for the growth of the parasite, however, remain largely unknown. Culture media for P. falciparum require human serum, a growth-promoting fraction derived from adult bovine plasma (GFS), or lipid-enriched bovine albumin (Asahi & Kanazawa, 1994;Asahi et al, 1996;Cranmer et al 1997;Jensen, 1979). Elucidation of the factors able to induce the growth of P. falciparum could be of help, not only for successful culture of the parasite, but also for providing critical clues to understanding the biology of parasite proliferation during the erythrocytic phase.…”

Section: Introductionmentioning

confidence: 99%

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Intraerythrocytic Plasmodium falciparum Growth in Serum-Free Medium with an Emphasis on Growth-Promoting Factors

Asahi¹

2012

Malaria Parasites

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“…To avoid the obstacles associated with the dependence on human serum, including unpredictable risks with infectious agents and difficulties in securing normal human sera, particularly in malaria endemic areas, and in obtaining a particular blood group, human serum substitutes such as a growth-promoting fraction derived from adult bovine plasma (GFS; Wako Pure Chemical Industries Ltd., Japan) and lipid-enriched bovine albumin (AlbuMAX I; Life Technologies, Japan) have been exploited success-fully [4,5], and have been employed widely for in vitro cultivation of P. falciparum. However, sufficient information on the constituents of the serum substitutes is still lacking.…”

Section: Introductionmentioning

confidence: 99%

Comparative studies of serum-free media and detection techniques for <i>in vitro</i> drug sensitivity assessment of <i>Plasmodium falciparum</i>

Kwansa–Bentum¹,

Izumiyama²,

Kitamura³

et al. 2013

OJCD

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Malaria continues to be a devastating disease. In a previous study, we formulated a chemically defined culture medium that is able to sustain the complete intraerythrocytic growth of Plasmodium falciparum. We tested the feasibility of using the medium (CDRPMI) as well as human serum-free media enriched with commercially available human-serum substitutes (GFSRPMI and ALBRPMI) to assess the drug sensitivity of P. falciparum, using chloroquine diphosphate (CQ) and dihydroartemisinin (DHART) as conventional antimalarial drugs. Growth inhibition was measured by four different methods: flow cytometry with SYBR Green I (FCM), microscopy (Giemsa method), enzymatic estimation of parasite lactate dehydrogenase (pLDH), and histidine-rich protein 2 (HRPII) determination. In drug sensitivity tests on asynchronous parasites cultured for 96 h in the presence of drugs, the dose-response curves were similar and differences in the 50% growth inhibition concentrations for the drugs, which were estimated by the four methods, were not statistically significant for the three culture media. The effect of the drugs on the growth of synchronous parasites at the ring stage was also assessed in micro-volume tests by three different methods of FCM: tracking fluorescent erythrocytes, schizont test, and merozoite test. Dose-response curves for the drugs were similar, and differences in the 50% growth inhibition concentrations were not statistically significant for CDRPMI and GFSRPMI.Thus CDRPMI as well as GFSRPMI and ALBRPMI can be similarly useful media for drug sensitivity testing of P. falciparum. The FCM, pLDH and HRPII estimations were fast and reliable detection methods, with FCM allowing schizont and merozoite tests to be performed with shorter periods of culture.

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An alternative to serum for cultivation of Plasmodium falciparum in vitro

Cited by 193 publications

References 15 publications

Malaria hemozoin is immunologically inert but radically enhances innate responses by presenting malaria DNA to Toll-like receptor 9

Malaria hemozoin is immunologically inert but radically enhances innate responses by presenting malaria DNA to Toll-like receptor 9

Intraerythrocytic Plasmodium falciparum Growth in Serum-Free Medium with an Emphasis on Growth-Promoting Factors

Comparative studies of serum-free media and detection techniques for <i>in vitro</i> drug sensitivity assessment of <i>Plasmodium falciparum</i>

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An alternative to serum for cultivation of Plasmodium falciparum in vitro

Cited by 193 publications

References 15 publications

Malaria hemozoin is immunologically inert but radically enhances innate responses by presenting malaria DNA to Toll-like receptor 9

Malaria hemozoin is immunologically inert but radically enhances innate responses by presenting malaria DNA to Toll-like receptor 9

Intraerythrocytic Plasmodium falciparum Growth in Serum-Free Medium with an Emphasis on Growth-Promoting Factors

Comparative studies of serum-free media and detection techniques for &lt;i&gt;in vitro&lt;/i&gt; drug sensitivity assessment of &lt;i&gt;Plasmodium falciparum&lt;/i&gt;

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Comparative studies of serum-free media and detection techniques for <i>in vitro</i> drug sensitivity assessment of <i>Plasmodium falciparum</i>