Trypanosoma vivax: a simplified protocol for in vivo growth, isolation and cryopreservation.
A rodent adapted clone of Trypanosoma vivax was used to infect cyclophosphamide treated mice and rats. Fresh blood containing trypanosomes, was centrifuged in a density gradient of three Percoll solutions, 1.07, 1.06, 1.05 g/ml, respectively, carefully layered on top of each other. The yields of this simple procedure for trypanosome purification were about six times higher than those obtained with the conventional anion-exchange columns. Cryopreservation of trypanosomes using glycerol yielded 90% viable parasites, whereas using dimethylsulfoxide, a more commonly used cryoprotectant, the viability was only 35%.
Ndao
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Institute of Tropical Medicine, Department of Parasitology, Nationalestraat 155, B-200 Antwerp, Belgium. momar.ndao@staff.mcgill.ca
Magnus
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Büscher
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Geerts
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D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient / Q000662:veterinary; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005260:Female; D005990:Glycerol / Q000494:pharmacology; D051379:Mice; D018512:Parasitemia / Q000469:parasitology / Q000662:veterinary; D051381:Rats; D017208:Rats, Wistar; D016019:Survival Analysis; D016689:Trypanosoma vivax / Q000187:drug effects / Q000254:growth & development* / Q000302:isolation & purification*; D014353:Trypanosomiasis, African / Q000469:parasitology / Q000662:veterinary*
D003451:Cryoprotective Agents; D005990:Glycerol; D004121:Dimethyl Sulfoxide