Affinity purification of Plasmodium ookinetes from in vitro cultures using extracellular matrix gel. (February 2021)

• Record Type: Journal Article
• Authors: Recio-Tótoro, Benito; Condé, Renaud; Claudio-Piedras, Fabiola; Lanz-Mendoza, Humberto
• Is Part Of: Parasitology international. Volume 80(2021)
• Publication Date: 2021-02
• Subjects: Plasmodium -- Ookinete -- Affinity purification -- Extracellular matrix gel; Parasitology -- Periodicals; Parasites -- Periodicals; Parasitic Diseases -- Periodicals; Parasitology -- Periodicals; Parasitologie -- Périodiques; 571.99905
• DOI: 10.1016/j.parint.2020.102242
• Languages: English
• ISSNs: 1383-5769

Abstract

Abstract: Malaria transmission depends on the parasites' successful invasion of the mosquito. This is achieved by the ookinete, a motile zygote that forms in the blood bolus after the mosquito takes an infectious blood meal. The ookinete invades the midgut epithelium and strongly attaches to the basal lamina, differentiating into an oocyst that produces the vertebrate-invasive sporozoites. Despite their importance, the ookinete and the oocyst are the least studied stages of the parasite. Much of what we know about the ookinete comes from in vitro experiments, which are hindered by the concomitant contamination with blood cells and other parasite stages. Although methods to purify them exist, they vary in terms of yield, costs, and difficulty to perform. A method for ookinete purification taking advantage of their adhesive properties was herein developed. The method consists of covering any culture-suitable surface with extracellular matrix gel, after which the ookinete culture is incubated on the gel to allow for ookinete attachment. The contaminant cells are then simply washed away. This procedure results in purer and less stressed ookinete preparations, which, by the nature of the method, are ready for oocyst production. Furthermore, it allows for micro-purifications using only 1 μl of blood, opening the possibility to make axenic ookinete cultures without sacrificing mice Graphical abstract: Unlabelled Image Highlights: In vitro Plasmodium ookinetes can be easily purified using an extracellular matrix gel. Ookinete selective binding allows their purification by rinsing the contaminant cells. This method is gentler to the ookinetes than the most common purification method. Ookinetes are ready for oocyst culture after purification with purities above 85%. Micro-cultures and micro-purifications can be performed without mice sacrifice.