Abstract
A simple technique for achieving high yields of Plasmodium falciparum parasites on a continuous basis is described. The technique is applicable in any laboratory. The culture apparatus is also simple and inexpensive and allows multiple cultures to be run simultaneously. A total of approximately 1-2 × 109 parasites can be harvested per culture flask per week requiring the use of only 40.0 ml of culture medium (RPMI 1640), 5.0 ml of human sera, and 2.0 ml of outdated human whole blood. P. falciparum parasites (segmenters containing individual merozoites) are cultured in vitro and concentrated 10-15 fold through the use of discontinuous bovine serum albumin gradient centrifugation.
Commercial saponin is purified on a Sephadex G-25 column. The haemolytic effect of purified saponin related to human red blood cell concentration is studied. Preliminary observations on the action of some synthetic detergents and enzymes on human erythrocytes are also reported. Purified saponin is used to lyse red blood cells infected with in vitro cultured P. falciparum for the preparation of merozoite antigen. Further purification of parasite material is carried out by sucrose density gradient centrifugation.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beach D. H., Sherman I. W., Holz G. G., Jr Lipids of Plasmodium lophurae, and of erythrocytes and plasma of normal and P. lophurae-infected Pekin ducklings. J Parasitol. 1977 Feb;63(1):62–75. [PubMed] [Google Scholar]
- Helenius A., Simons K. Solubilization of membranes by detergents. Biochim Biophys Acta. 1975 Mar 25;415(1):29–79. doi: 10.1016/0304-4157(75)90016-7. [DOI] [PubMed] [Google Scholar]
- Jensen J. B., Trager W. Plasmodium falciparum in culture: use of outdated erthrocytes and description of the candle jar method. J Parasitol. 1977 Oct;63(5):883–886. [PubMed] [Google Scholar]
- Siddiqui W. A. An effective immunization of experimental monkeys against a human malaria parasite, Plasmodium falciparum. Science. 1977 Jul 22;197(4301):388–389. doi: 10.1126/science.406671. [DOI] [PubMed] [Google Scholar]
- Siddiqui W. A., Kramer K., Richard-Crum S. M. In vitro cultivation and partial purification of Plasmodium falciparum antigen suitable for vaccination studies in Aotus monkeys. J Parasitol. 1978 Feb;64(1):168–169. [PubMed] [Google Scholar]
- Trager W., Jensen J. B. Human malaria parasites in continuous culture. Science. 1976 Aug 20;193(4254):673–675. doi: 10.1126/science.781840. [DOI] [PubMed] [Google Scholar]
- WARREN L. The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959 Aug;234(8):1971–1975. [PubMed] [Google Scholar]