The Development of the Malaria Parasite Plasmodium lophurae in Red Blood Cell Suspensions In vitro

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Abstract
Multiplication of P. lophurae has been obtained in suspensions of duck erythro-cytes maintained in vitro. Each 50-ml. culture flask is prepared to contain 4.5 ml. of culture medium and 1.5 ml. of a mixture of normal and parasitized duck blood such that the final suspension contains about 50,000 parasites and 1,000,000 red blood cells per cu. mm. The flask is equipped with gas inlet and outlet tubes through which is passed a slow current of moist 95% air-5%CO2. It is rocked gently on a rocking machine in an incubator at 39.5-40.5[degree]C. The culture me-dium consists of either a high-potassium red cell extract soln. previously used in survival studies with P. lophurae (Jour. Exptl. Med. 77: 411. 1943) or a medium developed for P. knowlesi (ibid. 84: 607. 1946) and containing vitamins, amino acids, purines and pyrimidines. The parasites multiply 2 to 3 times during a 2-day period of cultivation. If they are then subcultured to a fresh suspension of duck erythrocytes in fresh culture medium, they continue to multiply for a further 2 days. In this manner they have been kept in vitro for 8 days, at the end of which time they were still multiplying actively. Omission of vitamins from the culture medium was deleterious to the growth of the parasites, but biotin concs. ranging from 0.6 to 120 per ml. had no effect on the extent of multiplication.

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