Methodology

Effect of the haematocrit layer geometry on Plasmodium falciparum static thin-layer in vitro cultures

Jordi Ferrer^{* 1} , Marina D Rosal² , Jaume M Vidal^{* 2} , Clara Prats¹ , Joaquim Valls¹ , Esperanza A Herreros² , Daniel López¹ and Domingo Gargallo²

¹Escola Superior d'Agricultura de Barcelona, Campus del Baix Llobregat, Departament de Física i Enginyeria Nuclear, Universitat Politècnica de Catalunya, Avda. del Canal Olímpic 15, 08860 Castelldefels, Spain

²Drug Discovery Biology Group, Diseases of the Developing World Center, of GlaxoSmithKline R&D, Tres Cantos, Madrid, Spain

author email corresponding author email* Contributed equally

Malaria Journal 2008, 7:203doi:10.1186/1475-2875-7-203

Published:	8 October 2008

Abstract

Background

In vitro cultivation of Plasmodium falciparum is usually carried out through the continuous preservation of infected erythrocytes deposited in static thin layers of settled haematocrit. This technique, called the candle-jar method, was first achieved by Trager and Jensen in 1976 and has undergone slight modifications since then. However, no systematic studies concerning the geometry of the haematocrit layer have been carried out. In this work, a thorough investigation of the effects of the geometric culturing conditions on the parasite's development is presented.

Methods

Several experimental trials exploring different settings have been carried out, covering haematocrit layer depths that ranged from 6 mm to 3 mm and separation between the walls of the culturing device that ranged from 7.5 mm to 9 mm. The obtained results have been analysed and compared to different system-level models and to an Individual-Based Model.

Conclusion

In line with the results, a mechanism governing the propagation of the infection which limits it to the vicinity of the interface between the haematocrit layer and the culture medium is deduced, and the most appropriate configurations are proposed for further experimental assays.