The Use of Feeder Cells in the Cultivation of the Asexual Erythrocytic Stages of Plasmodium falciparum

Trenholme, K. R (1987) The Use of Feeder Cells in the Cultivation of the Asexual Erythrocytic Stages of Plasmodium falciparum. PhD thesis, University of Glasgow.

Abstract

While the technique of Trager and Jensen (1976) allows the in vitro cultivation of the asexual erythrocytic stages of P. falciparum, this method does have a number of limitations. Most isolates of P. falciparum grow poorly during the first weeks of cultivation and only some new isolates can eventually be established in culture. Methods to increase the proportion of isolates, and also the number of individual parasites within an isolate which could be established in long term culture were studied. Cultures of P. falciparum showed increased multiplication rates in the presence of a feeder cell layer of mouse peritoneal wash cells and within this population the adherent (macrophage enriched) population was the most important in promoting this increase. New isolates of P. falciparum adapted to culture more readily in the presence of a feeder cell layer of PWCS. Thirteen isolates were tested and only 3 were established in culture in the absence of PWCS. A further 6, however were adapted when cultured with PWCS. Three of the 4 isolates which were not established in culture with PWCS were known to have been taken from patients who had been treated with antimalarials and therefore the viability of these parasites at the time of culture was questionable. This was confirmed by the fact that these parasites did not complete a single asexual cycle. The effect of PWCS in the adaptation of new isolates of P. falciparum to long term culture was examined. It was found that the presence of PWCS led to the preservation of a greater degree of parasite diversity than that which was achieved under standard culture conditions. The presence of a feeder cell layer of mouse PWCS when cloning new isolates of P. falciparum (by limiting dilution) led to greatly increased cloning efficiency. Sixteen clones were produced from isolate AF using this method. These clones were examined using a panel of monoclonal antibodies and three different antigenic types were observed. In an attempt to streamline this cloning technique the feasibility of using cryopreserved PWCS was investigated. This was not, however, successful as the ability of cryopreserved PWCS to promote an increase in parasite growth declined rapidly during storage at -70

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Keywords: Parasitology
• Date of Award: 1987
• Depositing User: Enlighten Team
• Unique ID: glathesis:1987-77609
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 14 Jan 2020 11:53
• Last Modified: 14 Jan 2020 11:53
• URI: https://theses.gla.ac.uk/id/eprint/77609