Comparison of the effects of glycerol, dimethyl sulfoxide, and hydroxyethyl starch solutions for cryopreservation of avian red blood cells

Graham, Jennifer E.; Meola, Dawn M.; Kini, Nisha R; Hoffman, Andrew M.

doi:10.2460/ajvr.76.6.487

Cited by 18 publications

(12 citation statements)

References 14 publications

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“…The authors speculated that the low recovery rate could be due to small details in their protocol, including the addition rate of each aliquot of glycerol, time allowed for equilibration of glycerol with the RBCs, freezing rate of the samples or the deglycerolization process. 25 Another issue with glycerol cryopreservation encountered in our study was the unexpected agglutination during the glycerol wash steps in the feline samples. To rule out agglutination caused by removal of the anticoagulant during the wash steps, we added back anticoagulant to unit #4 both before the first washing steps (aliquot 2) and after the second wash (aliquot 3).…”

Section: Discussionmentioning

confidence: 89%

“…18 A manual technique for glycerolization and deglycerolization was also employed in a study of cryopreservation of avian red blood cells. 25 The avian study reported a recovery rate of 0% after the washing steps. The authors speculated that the low recovery rate could be due to small details in their protocol, including the addition rate of each aliquot of glycerol, time allowed for equilibration of glycerol with the RBCs, freezing rate of the samples or the deglycerolization process.…”

Section: Discussionmentioning

confidence: 99%

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Cryopreservation of feline red blood cells in liquid nitrogen using glycerol and hydroxyethyl starch

Hon

Thomovsky

Brooks

et al. 2019

Journal of Feline Medicine and Surgery

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Objectives The objective of this study was to evaluate the techniques and short-term effects of cryopreservation of feline red blood cells (RBCs) in liquid nitrogen using glycerol or hydroxyethyl starch (HES) as a cryoprotectant. Methods Feline RBCs were manually mixed with either 20% glycerol or 12.5% HES and frozen for 24 h in liquid nitrogen. The samples were thawed and glycerolized samples were manually washed. Success of the freeze/thaw process was determined by recovery rate of RBCs and evaluation of morphological changes using scanning electron microscopy (SEM). A unit of canine packed RBCs was also subjected to the same methodology to evaluate the cryopreservation handling technique. Results Feline RBCs preserved with 20% glycerol had a high recovery rate (94.23 ± 1.25%) immediately after thawing. However, the majority of the cells were lost during the washing process, with a final packed cell volume of <1%. A recovery rate was unable to be assessed for samples preserved with HES owing to the high viscosity of the mixture. SEM revealed significant morphological changes after glycerol was added to the feline RBCs. Although these morphological changes were partially reversed after thawing, the majority of the RBCs were lost during the washing process. Minimal morphological changes were noted in the HES sample. Similar results were noted with the canine RBCs. Conclusions and relevance The described manual technique for cryopreservation using glycerol was not able to successfully preserve feline or canine RBCs. In the present study, it was difficult to make conclusions about the efficacy of HES. Further studies evaluating HES as a cryoprotectant are warranted.

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Section: Discussionmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Cryopreservation of feline red blood cells in liquid nitrogen using glycerol and hydroxyethyl starch

Hon

Thomovsky

Brooks

et al. 2019

Journal of Feline Medicine and Surgery

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“…After sampling, six aliquots of whole blood (25 μl) per bird were created and 75 μl of freezing media (10% dimethyl sulfoxide [DMSO], 60% fetal bovine serum, and 30% modified Eagle medium‐α; Graham, Meola, Kini, & Hoffman, 2015) was added to each tube. Note that there are many cryopreservation methods and the one used here was based on one previously published method of avian red blood cell preservation (Graham et al, 2015). Three aliquots were then stored at −20°C and the other three aliquots were first slowly frozen to −80°C by surrounding the vials with isopropyl alcohol enclosed in a Styrofoam container overnight, then transferred to LN2.…”

Section: Methodsmentioning

confidence: 99%

Beyond corticosterone: The acute stress response increases DNA damage in house sparrows

et al. 2020

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Although corticosterone (Cort) has been the predominant metric used to assess acute stress in birds, it does not always accurately reflect how an animal copes with a stressor. Downstream measurements may be more reliable. In the current study, we tested the hypothesis that acute increases in DNA damage could be used to assess stressor exposure. Studies have shown DNA damage increases in response to stress‐related hormones in vitro; however, this has not yet been thoroughly applied in wild animals. We exposed house sparrows (Passer domesticus) to a 30‐ or 120‐min restraint stressor and took blood samples at 0, 30, 60, and 120 min to measure Cort, DNA damage, and uric acid. Both treatments increased DNA damage and Cort, and decreased uric acid. It thus appears that DNA damage can reflect acute stressor exposure. To improve the usability of DNA damage as a metric for stress, we also tested the impacts of sample storage on DNA damage. Leaving red blood cells on ice for up to 24 hr, only slightly influenced DNA damage. Freezing blood samples for 1–4 weeks substantially increased DNA damage. These findings emphasize the importance of reducing variation between samples by assaying them together whenever possible. Overall, these results indicate that assessing DNA damage is a valid method of assessing acute stressor exposure that is suitable for both laboratory‐ and field‐based studies; however, additional research is needed on the molecular dynamics of nucleated red blood cells, including whether and how their DNA is repaired.

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“…The advantages of cryopreservation methods with non-penetrating cryoprotectants for RBCs include the possibility to eliminate the washing steps prior to transfusion. However, the drawback of such CPAs is the presence of latent damage in thawed erythrocytes, which manifests itself in lowering the osmotic stability when they are placed in isotonic medium [24,25]. …”

Section: Introductionmentioning

confidence: 99%

Exploring the Possibility of Cryopreservation of Feline and Canine Erythrocytes by Rapid Freezing with Penetrating and Non-Penetrating Cryoprotectants

et al. 2017

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Efficient application of veterinary blood transfusion approaches for small companion animals requires readily available supply of the donor material. This can be achieved by developing of effective biobanking technologies allowing long-term storage of donor blood components via cryopreservation. Transfusion of an erythrocyte concentrate allows the successful correction of various hematological pathologies, severe bleeding, and etc. While in the past there were several approaches to cryopreserve red blood cells of dogs, to our knowledge there is virtually no data on cryopreservation of feline erythrocytes. In this paper, we performed a comprehensive parameter optimization for low temperature storage of RBCs of both species. Here, the efficiency of single-component and multicomponent cryoprotective media as well as necessary time of pre-incubation with penetrating and non-penetrating cryoprotectants prior to rapid freezing is analyzed. This study showed that glycerol was not sufficient for cryopreservation of red blood cells of the studied species under the investigated conditions. Application of 10% (v/v) ME2SO allowed for a significant reduction of canine and feline erythrocytes hemolysis after thawing. 17.5% hydroxyethyl starch demonstrated the highest cryoprotective activity for both species. It was found that dog RBCs should be incubated in cryoprotective media for 30 min at 22°C prior to freezing, while for cat RBCs 20 min is sufficient. Combination of CPAs was less effective. Presented data may be considered in further studies in veterinary transfusion and blood banking optimization.

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Comparison of the effects of glycerol, dimethyl sulfoxide, and hydroxyethyl starch solutions for cryopreservation of avian red blood cells

Abstract: Cryopreservation of avian RBCs with HES solution, regardless of HES concentration, resulted in greater degrees of apoptosis and cell death than did cryopreservation with other media. Transfusion with RBCs cryopreserved in HES solution may result in posttransfusion hemolysis in birds.

Cited by 18 publications

References 14 publications

Cryopreservation of feline red blood cells in liquid nitrogen using glycerol and hydroxyethyl starch

Cryopreservation of feline red blood cells in liquid nitrogen using glycerol and hydroxyethyl starch

Beyond corticosterone: The acute stress response increases DNA damage in house sparrows

Exploring the Possibility of Cryopreservation of Feline and Canine Erythrocytes by Rapid Freezing with Penetrating and Non-Penetrating Cryoprotectants

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