Hypoxanthine: a low molecular weight factor essential for growth of erythrocytic Plasmodium falciparum in a serum-free medium

A low molecular weight factor in a basal medium essential for erythrocytic Plasmodium falciparum development in a serum-free medium using a cell growth-promoting factor derived from adult bovine serum was detected. The factor was hypoxanthine. The optimal hypoxanthine concentration for parasite growth was between 15 and 120 microM. The contribution of hypoxanthine to increased parasite growth was clearly evident in cultures on day 4. Among various low molecular weight supplements tested, adenine, adenosine, AMP, ATP, cyclic AMP, guanine, guanosine, inosine, inosine monophosphate, xanthine, NAD, NADH, NADP, NADPH and deoxyguanosine triphosphate showed a similar effect to that of hypoxanthine in the serum-free culture system. On the other hand, the addition of uric acid, FAD, thymidine, uridine, orotic acid, deoxythymidine triphosphate, deoxycytidine triphosphate, deoxyadenosine triphosphate, ribose-1-phosphate, or ethanolamine was not beneficial to the parasite growth. The results presented here will not only be of practical value, but will provide important information about the developmental requirements of the parasite.

[1]  J. Jensen,et al.  Isolation and cultivation of Plasmodium falciparum using adult bovine serum. , 1985, The Journal of parasitology.

[2]  A. Sabin,et al.  The hemagglutinin of St. Louis encephalitis virus. I. Recovery of stable hemagglutinin from the brains of infected mice. , 1953, Journal of immunology.

[3]  I. W. Sherman Transport of amino acids and nucleic acid precursors in malarial parasites. , 1977, Bulletin of the World Health Organization.

[4]  H. Vial,et al.  Use of radioactive ethanolamine incorporation into phospholipids to assess in vitro antimalarial activity by the semiautomated microdilution technique , 1992, Antimicrobial Agents and Chemotherapy.

[5]  W. Gutteridge,et al.  Incorporation of radioactive precursors into DNA and RNA of Plasmodium knowlesi in vitro. , 1970, The Journal of protozoology.

[6]  I. Sherman,et al.  Biochemistry of Plasmodium (malarial parasites). , 1979, Microbiological reviews.

[7]  V. Clavey,et al.  Lipid traffic between high density lipoproteins and Plasmodium falciparum-infected red blood cells , 1991, The Journal of cell biology.

[8]  W. Trager,et al.  Human malaria parasites in continuous culture. , 1976, Science.

[9]  T. Kudo,et al.  A great improvement of fusion efficiency in mouse B cell hybridoma production by use of the new culture medium, GIT. , 1987, The Tohoku journal of experimental medicine.

[10]  H. Asahi,et al.  Continuous cultivation of intraerythrocytic Plasmodium falciparum in a serum-free medium with the use of a growth-promoting factor , 1994, Parasitology.

[11]  P. Rathod,et al.  Enzymes of purine and pyrimidine metabolism from the human malaria parasite, Plasmodium falciparum. , 1982, Molecular and biochemical parasitology.

[12]  A. Macleod,et al.  Plasmodium falciparum: modifications of the in vitro culture conditions improving parasitic yields. , 1982, The Journal of parasitology.

[13]  Gero Am,et al.  Purines and pyrimidines in malarial parasites. , 1990 .

[14]  C. Roberts,et al.  Cultivation of Plasmodium falciparum parasites in a serum-free medium. , 1993, The American journal of tropical medicine and hygiene.

[15]  G. Kugler A column chromatographic method for determination of plasma and erythrocyte levels of inosine and hypoxanthine. , 1978, Analytical biochemistry.

[16]  J. Jensen,et al.  Studies on serum requirements for the cultivation of Plasmodium falciparum. 2. Medium enrichment. , 1982, Bulletin of the World Health Organization.