Cryopreservation of Erythrocytes, Thrombocytes, and Lymphocytes

Sputtek, Andreas; Kühnl, P.; Rowe, Arthur W.

doi:10.1159/000104136

Cited by 25 publications

(26 citation statements)

References 32 publications

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“…However, granulocytes have been shown to be highly susceptible to damage during the cryopreservation process [3,5,41], and previous attempts at granulocyte cryopreservation have produced variable but largely unfavorable results [7]. At present, a clinically suitable method for granulocyte cryopreservation does not appear to be available [40]. …”

Section: Introductionmentioning

confidence: 99%

Membrane permeability of the human granulocyte to water, dimethyl sulfoxide, glycerol, propylene glycol and ethylene glycol

Vian

Higgins

2014

Cryobiology

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Granulocytes are currently transfused as soon as possible after collection because they rapidly deteriorate after being removed from the body. This short shelf life complicates the logistics of granulocyte collection, banking and safety testing. Cryopreservation has the potential to significantly increase shelf life; however, cryopreservation of granulocytes has proven to be difficult. In this study, we investigate the membrane permeability properties of human granulocytes, with the ultimate goal of using membrane transport modeling to facilitate development of improved cryopreservation methods. We first measured the equilibrium volume of human granulocytes in a range of hypo- and hypertonic solutions and fit the resulting data using a Boyle-van't Hoff model. This yielded an isotonic cell volume of 378 μm3 and an osmotically inactive volume of 165 μm3. To determine the permeability of the granulocyte membrane to water and cryoprotectant (CPA), cells were injected into well-mixed CPA solution while collecting volume measurements using a Coulter Counter. These experiments were performed at temperatures ranging from 4 to 37 °C for exposure to dimethyl sulfoxide, glycerol, ethylene glycol and propylene glycol. The best-fit water permeability was similar in the presence of all of the CPAs, with an average value at 21 °C of 0.18 μm atm−1 min−1. The activation energy for water transport ranged from 41 to 61 kJ/mol. The CPA permeability at 21 °C was 6.4, 1.0, 8.4 and 4.0 μm/min for dimethyl sulfoxide, glycerol, ethylene glycol and propylene glycol, respectively, and the activation energy for CPA transport ranged between 59 and 68 kJ/mol.

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Section: Introductionmentioning

confidence: 99%

Membrane permeability of the human granulocyte to water, dimethyl sulfoxide, glycerol, propylene glycol and ethylene glycol

Vian

Higgins

2014

Cryobiology

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“…Although the reasons for activity of PVA remain unclear at present, the ongoing search for other polymeric IRIs has not identified many active candidates 31,36 . Other non-penetrative polymers, poly (vinylpyrrolidone) and dextran, have also been explored as cryopreservatives but have minimal IRI activity 37,38 .…”

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confidence: 99%

Synthetic polymers enable non-vitreous cellular cryopreservation by reducing ice crystal growth during thawing

et al. 2014

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The cryopreservation of cells, tissue and organs is fundamental to modern biotechnology, transplantation medicine and chemical biology. The current state-of-the-art method of cryopreservation is the addition of large amounts of organic solvents such as glycerol or dimethyl sulfoxide, to promote vitrification and prevent ice formation. Here we employ a synthetic, biomimetic, polymer, which is capable of slowing the growth of ice crystals in a manner similar to antifreeze (glyco)proteins to enhance the cryopreservation of sheep and human red blood cells. We find that only 0.1 wt% of the polymer is required to attain significant cell recovery post freezing, compared with over 20 wt% required for solvent-based strategies. These results demonstrate that synthetic antifreeze (glyco)protein mimics could have a crucial role in modern regenerative medicine to improve the storage and distribution of biological material for transplantation.

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“…Cryoprotectants protect against cold shock damage due to freezing, intracellular crystallisation and de-crystallisation as well as membrane destabilisation during the thawing process. Prior to freezing intracellular balance is sustained by the help of cryoprotectants (Palasz and Mapletopt 1996;Kim et al 2004;Sputtek et al 2007;Chaudhari 2009). …”

mentioning

confidence: 99%

Freezing and storage of leukodepleted erythrocyte suspensions

Bala¹,

Eraslan²,

Akyazı³

et al. 2016

Vet. Med. - Czech

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Studies on the frozen storage of human blood products have benefited veterinary transfusion medicine in recent years, but the long-term cryopreservation of canine red blood cells (RBCs) has not yet been thoroughly investigated. Further, no studies are available with respect to the frozen storage of leukocyte-depleted canine red blood cells (LD-RBCs). The objective of the current study was to investigate time-dependent effects of long-term frozen storage on leukocyte-depleted canine RBCs. Twelve healthy adult dogs meeting the criteria for blood transfusion were used in the study. Whole blood samples (450 ± 45 ml) collected from each dog were centrifuged for 5 min at 22 °C and 4200 × g in a cryogenic microcentrifuge and concentrated RBC (pRBC) suspensions were obtained. Leukocyte depletion was achieved by filtration (2.6 log 10 ). Then, the filtrated samples were prewashed three times in 0.9% NaCl solution and were allocated into three subgroups to be evaluated at three different time points (Day 0, Month 4 and Month 6). The samples for cryopreservation were subjected to glycerolisation and then stored at -80 °C for 4-and 6-month periods. At the end of this period pRBC units were thawed by manual agitation in a water bath maintained at 36-38 °C, centrifuged and then washed in a consecutive series of 12%, 1.6% and 0.9% of NaCl + 0.2 dextrose solutions. 2,3-Diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), supernatant haemoglobin (SupHb), sodium (Na + ) and potassium (K + ) levels, residual glycerol concentrations and haemograms of thawed and deglycerolised pRBC samples were evaluated together with those of Day 0. Sterility tests were performed on all samples for bacterial contamination. No statistically significant differences were noted except for Hct and SupHb levels. No bacterial contamination was noted in any of the samples on the basis of sterility tests. It was found that the described glycerolisation procedure could be a method of choice in the cryopreservation of leukocyte-depleted pRBCs (LD-pRBCs) since no negative effect was observed on the quality of the products and long-term frozen storage did not cause RBC destruction.

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Cryopreservation of Erythrocytes, Thrombocytes, and Lymphocytes

Cited by 25 publications

References 32 publications

Membrane permeability of the human granulocyte to water, dimethyl sulfoxide, glycerol, propylene glycol and ethylene glycol

Membrane permeability of the human granulocyte to water, dimethyl sulfoxide, glycerol, propylene glycol and ethylene glycol

Synthetic polymers enable non-vitreous cellular cryopreservation by reducing ice crystal growth during thawing

Freezing and storage of leukodepleted erythrocyte suspensions

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