Abstract
BackgroundIn 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.MethodsSeveral 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.ConclusionIn 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.
Highlights
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
Plasmodium falciparum cultures were carried out in complete culture medium (RPMI 1640 medium supplemented with 25 mM HEPES, 10% human serum and 0.15 mM Hypoxanthine)
Red blood cells supplied by the Spanish Red Cross were added at 5% of haematocrit value (H = 5 0 ± 0.4%)
Summary
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 . Continuous culture of viable Plasmodium falciparum schizogonic stages was first achieved soon after by means of cultures that maintained infected red blood cells (RBCs) settled in a thin deposit under controlled conditions (static cultures). Trager and Jensen developed two types of static cultures: a system with continuous medium replacement, and a system with discrete daily medium renewal, called the candle-jar method [2]. Static cultivation of infected RBCs prevails as the most widespread parasite reservoir for clinical and pharmacological purposes
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