Cryopreservation of <i>Trypanosoma cruzi</i> Bloodstream Forms*

Filardi, Leny S.; Brener, Z.

doi:10.1111/j.1550-7408.1975.tb05190.x

Cited by 25 publications

(9 citation statements)

References 6 publications

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“…Interestingly, BSF cells in vitro which were treated using flash freezing showed growth advantages. As a matter of fact, a similar phenomenon has been described in Trypanosoma cruzi (Filardi & Brener, 1975). These results might be explained by a longer exposure time to the CPA during slow cooling resulting in higher toxicity.…”

Section: Discussionsupporting

confidence: 73%

Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei

Wen

Lyu

et al. 2016

Acta Tropica

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Trehalose, a non-permeating cryoprotective agent (CPA), has been documented as less toxic and highly efficient at cryopreserving different kinds of cells or organisms. In the present study, trehalose was evaluated for its application in cryopreservation of both Trypanosoma brucei procyclic and bloodstream form cells. The cryopreservation efficiency was determined by the motility of trypanosomes after thawing, as well as a subsequent recovery and infectivity assessment. The viability of trypanosomes from cultivation that were frozen in a serial concentrations of trehalose showed similar results to classical CPAs of glycerol and DMSO. Nevertheless, trypanosomes cryopreserved in 0.2M trehalose showed the best growth characteristic during subsequent cultivation. In addition, CPA cocktails with trehalose and permeating CPA glycerol or DMSO were developed and evaluated. Interestingly, trypanosomes in host (mouse) blood cryopreserved in 0.4M trehalose plus 5% glycerol showed higher infectivity than those preserved in trehalose/DMSO cocktails as well as individually. Further investigations showed that, in comparison with slow freezing at -80°C, flash freezing in liquid nitrogen provided better cryopreservation for bloodstream form cells than slow freezing. In conclusion, trehalose is an easy, safe and efficient CPA for cryopreservation of T. brucei and potentially for other protozoan species and cells.

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Section: Discussionsupporting

confidence: 73%

Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei

Wen

Lyu

et al. 2016

Acta Tropica

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“…Blood protozoa, such as Trypanosoma spp. and Plasmodium spp., can be frozen easily without cryoprotectants or precise control of the freezing rates [4,8,14]. On the contrary, studies on the cryopreservation of helminths have been limited to only a few nematodes.…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation of protozoan parasites

2004

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Conventional methods for the propagation and preservation of parasites in vivo or in vitro have some limitations, including the need for labor, initial isolation and loss of strains, bacterial, and fungal contamination, and changes in the original biological and metabolic characteristics. All these disadvantages are considerably reduced by cryopreservation. In this study, we examined the effects of various freezing conditions on the survival of several protozoan parasites after cryopreservation. The viability of Entamoeba histolytica was improved by seeding (p < 0.05, chi2 test), while this was not so effective for Trichomonas vaginalis. Of six cryoprotectants examined, dimethyl sulfoxide (Me(2)SO), and glycerol showed the strongest cryoprotective effects. The optimum conditions for using Me(2)SO were a concentration of 10% with no equilibration, and those for glycerol were a concentration of 15% with equilibration for 2h. The optimum cooling rate depended on the parasite species. Trypanosoma brucei gambiense and Leishmania amazonensis were successfully cryopreserved over a wide range of cooling rates, whereas the survival rates of E. histolytica, T. vaginalis, Pentatrichomonas hominis, and Blastocystis hominis were remarkably decreased when frozen at improper rates. Unlike the cooling rate, exposure of the protozoans to a rapid thawing method produced better motility for all parasites.

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“…Special attention should be paid to nuclear changes, considering that condensation of the karyoplasm or even denaturation of chromatin does not seem to have an effect on further cultivation (Dar et al 1972;Filardi and Brener 1975). As these nuclear alterations were very fre-quently observed by means of TEM we would suggest that they are not lethal to trypanosomes -at least not to all of them -because the observed successful infections could not otherwise be understood.…”

Section: Discussionmentioning

confidence: 96%

Ultrastructural changes on various Trypanosoma spp. after a 30-year storage period in liquid nitrogen

Schuster

Mehlhorn

Raether³

1996

Parasitology Research

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Four Trypanosoma species were examined for damage following prolonged storage in liquid nitrogen (-196 degrees C). The stabilates were successfully recovered after a cryopreservation period of approximately 30 years. The structure of specimens was studied by means of light microscopy and scanning (SEM) and transmission (TEM) electron microscopy. All of the species tested--T. evansi, T. equinum, T. brucei, and T. congolense--proved to be infective to mice. However, as compared with controls, the trypomastigote bloodstream forms, which had been frozen and later recovered, showed clear differences. Formerly deep-frozen organisms usually appeared to have shrunk as a result of solution effects, which occur during freezing and thawing. Ultrastructural changes such as separation of the cytoplasm from the pellicle, the occurrence of large vacuoles in the cytoplasm and karyoplasm, a loss of cytoplasmatic ribosomes, membrane injuries, enlargement of the flagellar pocket, and denaturation of chromatin became obvious. The extent of the ultrastructural alterations appeared to be much greater after a cryopreservation period of approximately 30 years than those previously reported after a 13-year storage period. These changes, however, did not result in a complete loss of infectivity to mice.

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Cryopreservation of Trypanosoma cruzi Bloodstream Forms*

Cited by 25 publications

References 6 publications

Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei

Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei

Cryopreservation of protozoan parasites

Ultrastructural changes on various Trypanosoma spp. after a 30-year storage period in liquid nitrogen

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