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Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes

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 inves...

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Autores principales: Ribacke, Ulf, Moll, Kirsten, Albrecht, Letusa, Ahmed Ismail, Hodan, Normark, Johan, Flaberg, Emilie, Szekely, Laszlo, Hultenby, Kjell, Persson, Kristina E. M., Egwang, Thomas G., Wahlgren, Mats
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3718792/
https://www.ncbi.nlm.nih.gov/pubmed/23894537
http://dx.doi.org/10.1371/journal.pone.0069781
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author Ribacke, Ulf
Moll, Kirsten
Albrecht, Letusa
Ahmed Ismail, Hodan
Normark, Johan
Flaberg, Emilie
Szekely, Laszlo
Hultenby, Kjell
Persson, Kristina E. M.
Egwang, Thomas G.
Wahlgren, Mats
author_facet Ribacke, Ulf
Moll, Kirsten
Albrecht, Letusa
Ahmed Ismail, Hodan
Normark, Johan
Flaberg, Emilie
Szekely, Laszlo
Hultenby, Kjell
Persson, Kristina E. M.
Egwang, Thomas G.
Wahlgren, Mats
author_sort Ribacke, Ulf
collection PubMed
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% O(2), 5% CO(2), 90% N(2)). 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.
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spelling pubmed-37187922013-07-26 Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes Ribacke, Ulf Moll, Kirsten Albrecht, Letusa Ahmed Ismail, Hodan Normark, Johan Flaberg, Emilie Szekely, Laszlo Hultenby, Kjell Persson, Kristina E. M. Egwang, Thomas G. Wahlgren, Mats PLoS One Research Article 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% O(2), 5% CO(2), 90% N(2)). 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. Public Library of Science 2013-07-22 /pmc/articles/PMC3718792/ /pubmed/23894537 http://dx.doi.org/10.1371/journal.pone.0069781 Text en © 2013 Ribacke et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Ribacke, Ulf
Moll, Kirsten
Albrecht, Letusa
Ahmed Ismail, Hodan
Normark, Johan
Flaberg, Emilie
Szekely, Laszlo
Hultenby, Kjell
Persson, Kristina E. M.
Egwang, Thomas G.
Wahlgren, Mats
Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes
title Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes
title_full Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes
title_fullStr Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes
title_full_unstemmed Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes
title_short Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes
title_sort improved in vitro culture of plasmodium falciparum permits establishment of clinical isolates with preserved multiplication, invasion and rosetting phenotypes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3718792/
https://www.ncbi.nlm.nih.gov/pubmed/23894537
http://dx.doi.org/10.1371/journal.pone.0069781
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