131:
183:) significantly increased culture growth. Continuous agitation has also been shown to improve other parameters of culture growth relevant to researchers, such as the prolongation of culture synchrony after synchronization procedures, and a reduction of the rate of multiple infections. Despite this, the practice of culturing the parasite under static conditions remains widespread. The greatest value of the candlejar method is that it can be used in laboratories almost anywhere in the world where there is an incubator, a candle and a
20:
204:
1464:
215:
68:
46:
environment. Although attempts for propagation of the parasites outside of humans or animal models reach as far back as 1912, the success of the initial attempts was limited to one or just a few cycles. The first successful continuous culture was established in 1976. Initial hopes that the
256:
in infected red blood cells can also be used for isolation. Magnetic columns have shown to be less harmful for the parasite and are simple and adjustable to the needs of the researcher. The column is mounted in a potent magnet holder and the culture flowed through it. The column traps the
257:
erythrocytes infected with the latest stages of the parasites, which can then be eluted when the column is removed from the magnet. It is a simple method that does not need expensive equipment and it does not seem to affect the parasites as to their invasion capabilities afterwards.
174:, placed in a candlejar and incubated. The line grew very well and became FCR-3/Gambia, one of the most widely used strains. Later, other lines would be established using similar methods and the impact of continuous cultivation of
453:"Molecular epidemiology of malaria in Cameroon. XV. Experimental studies on serum substitutes and supplements and alternative culture media for in vitro drug sensitivity assays using fresh isolates of Plasmodium falciparum"
592:
Rüssmann L, Jung A, Heidrich HG (1982). "The use of percoll gradients, elutriator rotor elution, and mithramycin staining for the isolation and identification of intraerythrocytic stages of plasmodium berghei".
178:
was phenomenal especially for the testing of putative antimalarials and for deciphering its genes. A number of subsequent reports (from as far back as the early 1980s), showed that cell suspension (using a
1451:
1441:
279:
Radfar A, Méndez D, Moneriz C, Linares M, Marín-García P, Puyet A, Diez A, Bautista JM (2009). "Synchronous culture of
Plasmodium falciparum at high parasitemia levels".
99:. An alternative to gassing the cultures with the exact gas mixture, is the use of a candlejar. The candlejar is an airtight container in which the cultures and a lit
119:
concentration reaches approximately 5 %, the candle stops burning. The number of parasites increased by a factor 5 approximately every 48 hours (one cycle). The
241:
from 1.075 to 1.100 g/ml, whereas schizonts were concentrated at a density approximating 1.062 g/ml. There are studies, however, that suggest that some strains of
493:
735:"Comparison of the in vitro invasive capabilities of Plasmodium falciparum schizonts isolated by Percoll gradient or using magnetic based separation"
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isolated directly from infected patients indicate that alternative parasite biological states occur in the natural host that are not observed with
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procedure can be used to isolate infected red blood cells because red cells containing plasmodia are less dense than normal ones. Young
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incubator. In the summer of 1976 Milton
Friedman, a graduate student in the Trager lab who was working in the MRC laboratories in
150:
described culture of the parasite under static conditions (Trager-Jensen method). James B. Jensen joined Trager’s laboratory as a
1431:
1426:
695:
LeRoux M, Lakshmanan V, Daily JP (2009). "Plasmodium falciparum biology: analysis of in vitro versus in vivo growth conditions".
234:
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To achieve synchronization and/or concentration of the parasites in culture several methods have been developed. A discontinuous
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130:
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494:"Recent advances in malaria parasite cultivation and their application to studies on host-parasite relationships: a review"
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or serum substitutes. A special feature of the incubation is the special gas mixture filled with
Nitrogen (90-92 %) CO
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Bhakdi SC, Ottinger A, Somsri S, Sratogno P, Pannadaporn P, Chimma P, Malasit P, Pattanapanyasat K, Neumann HP (2010).
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Trager W, Jensen JB (1997). "Continuous culture of
Plasmodium falciparum: its impact on malaria research".
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are affected in their capacity of invasion after being exposed to this chemical. The difference between
187:. Around 60% parasitized cells can be obtained using optimized culturing conditions. Recent studies of
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127:, to keep it within the wanted limits, the culture can be thinned out with healthy red blood cells.
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928:"Optimized high gradient magnetic separation for isolation of Plasmodium-infected red blood cells"
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115:. Carbon dioxide content in fresh air varies between 0.036 % and 0.039 %. Once the CO
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835:"Separation and concentration of schizonts of Plasmodium falciparum by Percoll gradients"
786:"Separation of Plasmodium falciparum Late Stage-infected Erythrocytes by Magnetic Means"
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95:(5 %) and Oxygen (3-5 %) allowing the parasites to grow at 37 °C in a
319:"The Cultivation of Malarial Plasmodia (Plasmodium vivax and Plasmodium Falciparum)
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138:-infected erythrocytes by discontinuous density gradient centrifugation in Percoll.
84:
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874:"Improved methods for magnetic purification of malaria parasites and haemozoin"
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Trager W, Jensen JB (1976). "Human malaria parasites in continuous culture".
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are placed. The burning candle consumes some of the oxygen and produces
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are incubated in a culture dish or flask at 37 °C together with a
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34:
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Coronado LM, Tayler NM, Correa R, Giovani RM, Spadafora C (2013).
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18:
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984:
Malaria
Methods and Protocols (Methods in Molecular Medicine)
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European and
Developing Countries Clinical Trials Partnership
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coincided with erythrocytes in a broad band corresponding to
170:(which turned out to be the best of the commercial media) in
154:
in 1976. He decided to employ a candlejar instead of the CO
672:
Magic
Bullets to Conquer Malaria. From Quinine to Qinghaosu
1020:
249:
low-spin oxyhemoglobin in uninfected red blood cells and
1442:
The Global Fund to Fight AIDS, Tuberculosis and
Malaria
162:, arranged for a sample of human blood infected with
16:
Method for growing malaria parasites outside the body
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166:to be sent to New York City. This was diluted with
51:culture would lead quickly to the discovery of a
833:Rivadeneira EM, Wasserman M, Espinal CT (1983).
55:were premature. However, the development of new
733:Spadafora C, Gerena L, Kopydlowski KM (2011).
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142:The original method for the successful
872:Kim CC, Wilson EB, Derisi JL (2010).
7:
1347:National Malaria Eradication Program
1402:Bill & Melinda Gates Foundation
632:culture: The benefits of shaking".
851:10.1111/j.1550-7408.1983.tb02932.x
14:
1463:
1462:
1432:African Leaders Malaria Alliance
1427:South African Malaria Initiative
982:• Doolan, D. L. (Editor) (2002)
646:10.1016/j.molbiopara.2009.09.005
1285:Malaria antigen detection tests
545:"Cultivation of plasmodium spp"
199:Concentration of infected cells
1369:Malaria in Mandatory Palestine
40:outside the body, i.e., in an
1:
1447:Medicines for Malaria Venture
1422:African Malaria Network Trust
561:10.1128/CMR.15.3.355-364.2002
430:10.1016/S0020-7519(97)00080-5
1332:Millennium Development Goals
986:, Totowa, NJ: Humana Press,
1503:
1397:Against Malaria Foundation
628:Allen RJ, Kirk K (2010). "
1460:
1437:Amazon Malaria Initiative
470:10.4269/ajtmh.2003.69.168
59:was greatly facilitated.
1364:Malaria in the Caribbean
1213:Sterile insect technique
709:10.1016/j.pt.2009.07.005
501:Bull. World Health Organ
33:is a method for growing
1417:Africa Fighting Malaria
1263:Diagnosis and treatment
752:10.1186/1475-2875-10-96
634:Mol. Biochem. Parasitol
218:Magnetic collection of
1181:Control and prevention
948:10.1186/1475-2875-9-38
894:10.1186/1475-2875-9-17
395:10.1126/science.781840
293:10.1038/nprot.2009.198
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195:cultivated parasites.
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123:can be determined via
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1374:Malaria Atlas Project
630:Plasmodium falciparum
532:on November 28, 2003.
457:Am. J. Trop. Med. Hyg
217:
206:
133:
70:
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1270:Diagnosis of malaria
1086:Pregnancy-associated
549:Clin. Microbiol. Rev
543:Schuster FL (2002).
339:10.1084/jem.16.4.567
317:, Johns FM. (1912).
152:post-doctoral fellow
1327:Diseases of poverty
1320:Society and malaria
1230:Sickle-cell anaemia
1203:Malaria prophylaxis
387:1976Sci...193..673T
111:), which acts as a
1407:Imagine No Malaria
1392:Malaria Consortium
1337:History of malaria
1220:Genetic resistance
1145:Anopheles mosquito
607:10.1007/BF00925344
224:
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73:
28:
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1354:World Malaria Day
1000:978-0-89603-823-3
681:978-1-55581-543-1
492:Trigg PI (1985).
451:Basco LK (2003).
418:Int. J. Parasitol
381:(4254): 673–675.
134:Concentration of
113:fire extinguisher
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1299:Artemisinin
1235:Thalassemia
790:J. Vis. Exp
640:(1): 63–5.
327:J. Exp. Med
281:Nat. Protoc
247:diamagnetic
121:parasitemia
83:medium and
1304:Mefloquine
1280:Blood film
1170:Gametocyte
1165:Hypnozoite
1150:Life cycle
1116:falciparum
1103:life cycle
1093:Plasmodium
261:References
185:desiccator
160:The Gambia
125:blood film
1309:Proguanil
1160:Merozoite
1011:Protocols
941:(1): 38.
887:(1): 17.
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239:densities
181:incubator
168:RPMI 1640
71:Candlejar
38:parasites
1481:Category
1468:Category
1155:Schizont
1137:knowlesi
1130:malariae
1071:Cerebral
967:20122252
935:Malar. J
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881:Malar. J
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739:Malar. J
717:19747879
670:(2010).
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321:in vitro
301:20010926
254:hemozoin
231:gradient
179:shaking-
81:nutrient
26:cultures
1487:Malaria
1098:biology
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1059:Biology
1052:Malaria
1016:Percoll
958:2831011
904:2817699
859:6313915
811:3622091
762:3100259
615:6177116
523:3893779
514:2536397
438:9363481
383:Bibcode
375:Science
348:2124976
315:Bass CC
228:Percoll
193:ex vivo
144:ex vivo
53:vaccine
49:ex vivo
43:ex vivo
35:malaria
998:
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567:
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436:
403:781840
401:
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299:
101:candle
85:plasma
63:Method
1252:RTS,S
1123:ovale
1109:vivax
931:(PDF)
877:(PDF)
530:(PDF)
497:(PDF)
89:serum
57:drugs
996:ISBN
988:ISBN
963:PMID
909:PMID
855:PMID
816:PMID
767:PMID
713:PMID
676:ISBN
650:PMID
611:PMID
575:PMID
519:PMID
475:PMID
434:PMID
399:PMID
353:PMID
297:PMID
1193:DDT
953:PMC
943:doi
899:PMC
889:doi
847:doi
806:PMC
798:doi
757:PMC
747:doi
705:doi
642:doi
638:169
603:doi
565:PMC
557:doi
509:PMC
465:doi
426:doi
391:doi
379:193
343:PMC
335:doi
289:doi
107:(CO
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