Magnetic isolation of Plasmodium falciparum schizonts iRBCs to generate a high parasitaemia and synchronized in vitro culture

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• 作者：Lydia Mata-Cantero (1) (2)
  Maria J Lafuente (1)
  Laura Sanz (1)
  Manuel S Rodriguez (2) (3)
  
  1. Medicines Development Campus ; Diseases of the Developing World ; GlaxoSmithKline ; Severo Ochoa 2 ; Tres Cantos ; 28760 ; Madrid ; Spain
  2. Proteomics Unit ; CICbioGUNE Ed ; 801A Parque Tecnol贸gico de Bizkaia ; 48160 ; Derio ; Spain
  3. Ubiquitylation and Cancer Molecular Biology ; Inbiomed ; Mikeletegi 81 ; 20009 ; San Sebastian ; Spain
  
• 关键词：Plasmodium falciparum ; High parasitaemia ; Synchronous cultures ; Magnetic columns
• 刊名：Malaria Journal
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：13
• 期：1
• 全文大小：739 KB
• 参考文献：WHO Expert Committee on Malaria. World Health Organ Tech Rep Ser 892: pp. 1-74
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• 刊物主题：Parasitology; Infectious Diseases; Tropical Medicine;
• 出版者：BioMed Central
• ISSN：1475-2875

文摘

Background The establishment of methods for an in vitro continuous culture of Plasmodium falciparum is essential for gaining knowledge into its biology and for the development of new treatments. Previously, several techniques have been used to synchronize, enrich and concentrate P. falciparum, although obtaining cultures with high parasitaemia continues being a challenging process. Current methods produce high parasitaemia levels of synchronized P. falciparum cultures by frequent changes of culture medium or reducing the haematocrit. However, these methods are time consuming and sometimes lead to the loss of synchrony. Methods A procedure that combines Percoll and sorbitol treatments, the use of magnetic columns, and the optimization of the in vitro culture conditions to reach high parasitaemia levels for synchronized Plasmodium falciparum cultures is described. Results A new procedure has been established using P. falciparum 3D7, combining previous reported methodologies to achieve in vitro parasite cultures that reach parasitaemia up to 40% at any intra-erythrocytic stage. High parasitaemia levels are obtained only one day after magnetic column purification without compromising the parasite viability and synchrony. Conclusions The described procedure allows obtaining a large scale synchronized parasite culture at a high parasitaemia with less manipulations than other methods previously described.