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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes
PLoS ONE, Volume 8, No. 7, Article e69781, Year 2013
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Description
To be able to robustly propagate P. falciparum at optimal conditions in vitro is of fundamental importance for genotypic and phenotypic studies of both established and fresh clinical isolates. Cryo-preserved P. falciparum isolates from Ugandan children with severe or uncomplicated malaria were investigated for parasite phenotypes under different in vitro growth conditions or studied directly from the peripheral blood. The parasite cultures showed a minimal loss of parasite-mass and preserved percentage of multiple infected pRBCs to that in peripheral blood, maintained adhesive phenotypes and good outgrowth and multiplication rates when grown in suspension and supplemented with gas. In contrast, abnormal and greatly fluctuating levels of multiple infections were observed during static growth conditions and outgrowth and multiplication rates were inferior. Serum, as compared to Albumax, was found necessary for optimal presentation of PfEMP1 at the pRBC surface and/or for binding of serum proteins (immunoglobulins). Optimal in vitro growth conditions of P. falciparum therefore include orbital shaking (50 rev/min), human serum (10%) and a fixed gas composition (5% O2, 5% CO2, 90% N2). We subsequently established 100% of 76 frozen patient isolates and found rosetting with schizont pRBCs in every isolate (>26% schizont rosetting rate). Rosetting during schizogony was often followed by invasion of the bound RBC as seen by regular and time-lapse microscopy as well as transmission electron microscopy. The peripheral parasitemia, the level of rosetting and the rate of multiplication correlated positively to one another for individual isolates. Rosetting was also more frequent with trophozoite and schizont pRBCs of children with severe versus uncomplicated malaria (p<0.002; p<0.004). The associations suggest that rosetting enhances the ability of the parasite to multiply within the human host. © 2013 Ribacke et al.
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC3718792/bin/pone.0069781.s001.pdf
https://efashare.b-cdn.net/share/pmc/articles/PMC3718792/bin/pone.0069781.s002.tif
https://efashare.b-cdn.net/share/pmc/articles/PMC3718792/bin/pone.0069781.s003.pdf
https://efashare.b-cdn.net/share/pmc/articles/PMC3718792/bin/pone.0069781.s004.doc
https://efashare.b-cdn.net/share/pmc/articles/PMC3718792/bin/pone.0069781.s005.mov
Authors & Co-Authors
Ribacke, Ulf
Sweden, Stockholm
Karolinska Institutet
United States, Boston
Harvard T.h. Chan School of Public Health
Moll, Kirsten
Sweden, Stockholm
Karolinska Institutet
Albrecht, Letusa
Sweden, Stockholm
Karolinska Institutet
Brazil, Campinas
Universidade Estadual de Campinas
Ahmed Ismail, Hodan
Sweden, Stockholm
Karolinska Institutet
Normark, Johan
Sweden, Stockholm
Karolinska Institutet
Sweden, Umea
Umeå Universitet
Flaberg, Emilie
Sweden, Stockholm
Karolinska Institutet
Székely, Lászlõ
Sweden, Stockholm
Karolinska Institutet
Hultenby, Kjell
Sweden, Stockholm
Karolinska Institutet
Persson, Kristina E.M.
Sweden, Stockholm
Karolinska Institutet
Egwang, Thomas Gordon
Uganda, Kampala
Med Biotech Laboratories Uganda
Wahlgren, Mats
Sweden, Stockholm
Karolinska Institutet
Statistics
Citations: 11
Authors: 11
Affiliations: 5
Identifiers
Doi:
10.1371/journal.pone.0069781
e-ISSN:
19326203
Research Areas
Health System And Policy
Infectious Diseases
Maternal And Child Health