Cryopreservation of Plasmodium gallinaceum brumpt sporozoites for 16 years at −196 °C

Ab, Weathersby; Jw, McCall

doi:10.1016/0011-2240(81)90103-6

Cited by 4 publications

(4 citation statements)

References 10 publications

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“…[9], and T. canis [10]. For example, Weathersby and McCall [14] reported that Plasmodium gallinaceum frozen without cryoprotectants was as infective as the nonfrozen organism.…”

Section: Discussionmentioning

confidence: 99%

“…Babesia rodhaini [2], T. cruzi [4], and Plasmodium spp. [8,14] were successfully cryopreserved by this method. The slow freezing method uses a programming freezer or alcohol bath to control the freezing rate (1-20℃/min), and this method has been applied to protozoa such as T. gondii [1], E. histolytica [3], G. lamblia [7], and T. vaginalis [12].…”

Section: Discussionmentioning

confidence: 99%

“…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%

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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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“…[9], and T. canis [10]. For example, Weathersby and McCall [14] reported that Plasmodium gallinaceum frozen without cryoprotectants was as infective as the nonfrozen organism.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Cryopreservation of protozoan parasites

2004

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“…Cryopreserving irradiated P. berghei sporozoites in the presence of serum has been successful, also [21]. In a surprising contrast, cryopreservation of P. gallinaceum sporozoites did not use any cryogenic solution but preserved sporozoites in situ in infected Aedes aegypti [22].…”

Section: Discussionmentioning

confidence: 99%

A simple and efficient method for cryopreservation and recovery of viable Plasmodium vivax and P. falciparum sporozoites

Singh

Barnes

Jenwithisuk

et al. 2016

Parasitology International

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Plasmodium falciparum and Plasmodium vivax sporozoites are the crucial stages of malaria parasites that initiate infection in humans. However, studies to develop new vaccines and drugs targeting these infective stages remain insufficient due to limited availability of sporozoites for research. This is a consequence of relatively few facilities that are established to produce sporozoites of human malaria parasites, sporozoites remaining viable for only a few days, and infected mosquitoes being a biohazard, making them difficult to transport. Cryopreservation of sporozoites offers the potential to alleviate these limitations and enhance sporozoite availability. These experiments were performed to evaluate methods for cryopreservation of P. vivax and P. falciparum sporozoites. Sporozoites, isolated in sterile buffer from infected mosquitoes by manual dissection of salivary glands, were cryopreserved using several types of commercially available serum-free cryoprotective solutions. The efficiency of cryopreservation was validated by a standard in vitro gliding motility assay as a measure of sporozoite activity. Viability of infective sporozoites was defined as percent gliding of sporozoites attached to the coverslip. Significant differences were observed among the cryopreservation media and protocols evaluated, with CryoStor CS2 giving the best results for both P. falciparum and P. vivax, whereas Hestar 200 worked efficiently only for P. vivax sporozoites. Further improvement in recovery of viable sporozoites would be anticipated using automated controlled-rate freezing equipment. Our results demonstrate that cryopreservation provides an alternative for experimental studies that currently rely on fresh P. falciparum and P. vivax sporozoites.

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Visual Observation of African Trypanosomes During Cryopreservation

Wang

Wen²,

Lun³

et al. 2014

Biopreservation and Biobanking

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Protozoa have been widely used for the study of cryopreservation. The survival rate after cryopreservation has always received the most attention, while the cell viability during the process of freezing and thawing has been much less studied. In the present study, we report successful cryopreservation of Trypanosoma brucei, a parasitic protozoa of human and animals, using controlled-rate freezing at 5°C/min, and real-time observation of activity using a microscope differential scanning calorimeter system during the freezing and thawing process. Trehalose used as a cryoprotective agent at a concentration of 0.4 M allowed the trypanosomes to endure freezing and thawing with >89% survival rate. Results from mechanisms analysis indicate that vitrification by trehalose contributes significantly to the protection of the trypanosomes from damage at low temperature.

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Cryopreservation of Plasmodium gallinaceum brumpt sporozoites for 16 years at −196 °C

Cited by 4 publications

References 10 publications

Cryopreservation of protozoan parasites

Cryopreservation of protozoan parasites

A simple and efficient method for cryopreservation and recovery of viable Plasmodium vivax and P. falciparum sporozoites

Visual Observation of African Trypanosomes During Cryopreservation

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