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Updated: Aug 24, 2026
Ultrastructural Expansion Microscopy in Three In Vitro Life Cycle Stages of Trypanosoma cruzi
Published on: May 12, 2023
Trypanosoma vivax: a simplified protocol for in vivo growth, isolation and cryopreservation
1Institute of Tropical Medicine, Department of Parasitology, Nationalestraat 155, B-200 Antwerp, Belgium. momar.ndao@staff.mcgill.ca
Abstract:
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%.
Insights
This study introduces a novel, high-yield method for purifying Trypanosoma vivax parasites using density gradient centrifugation. Glycerol proved superior to dimethylsulfoxide for cryopreservation, ensuring high parasite viability.
Area of Science:
- Veterinary Parasitology
- Cell Biology
- Biotechnology
Background:
- Trypanosoma vivax is a significant veterinary pathogen.
- Efficient purification and cryopreservation methods are crucial for Trypanosoma research and control.
- Existing purification techniques can be labor-intensive and yield suboptimal results.
Purpose of the Study:
- To develop an improved method for purifying Trypanosoma vivax.
- To evaluate glycerol and dimethylsulfoxide as cryoprotectants for Trypanosoma vivax.
- To enhance parasite viability post-cryopreservation.
Main Methods:
- Infection of immunosuppressed rodents (mice and rats) with a rodent-adapted Trypanosoma vivax clone.
- Purification of trypanosomes from infected blood using a three-step Percoll density gradient centrifugation.
- Cryopreservation of purified trypanosomes using glycerol and dimethylsulfoxide.
Main Results:
- Density gradient centrifugation yielded approximately six times more trypanosomes compared to conventional anion-exchange columns.
- Cryopreservation with glycerol resulted in 90% parasite viability.
- Cryopreservation with dimethylsulfoxide yielded only 35% parasite viability.
Conclusions:
- Percoll density gradient centrifugation is a highly efficient method for purifying Trypanosoma vivax.
- Glycerol is a superior cryoprotectant for Trypanosoma vivax compared to dimethylsulfoxide.
- The developed method significantly improves trypanosome purification yields and cryopreservation success rates.
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