Membrane Stability during Biopreservation of Blood Cells

Stoll, Christoph; Wolkers, Willem F.

doi:10.1159/000326900

Cited by 31 publications

(19 citation statements)

References 69 publications

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“…The cellular membrane is prone to injury during rapid cooling and phase transitions (cryopreservation) [32,33] or prolonged cold exposures (CS) [4,34]. SEM imaging of SCP hepatocytes reveal that membrane injury is also present during supercooling preservation; however, whether this damage is the result of direct lipid peroxidation or phase transition or an internal cascade manifesting (apoptosis, ion imbalance) itself as eventual blebbing is unclear.…”

Section: Discussionmentioning

confidence: 99%

Supercooling as a Viable Non-Freezing Cell Preservation Method of Rat Hepatocytes

et al. 2013

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Supercooling preservation holds the potential to drastically extend the preservation time of organs, tissues and engineered tissue products, and fragile cell types that do not lend themselves well to cryopreservation or vitrification. Here, we investigate the effects of supercooling preservation (SCP at -4oC) on primary rat hepatocytes stored in cryovials and compare its success (high viability and good functional characteristics) to that of static cold storage (CS at +4oC) and cryopreservation. We consider two prominent preservation solutions a) Hypothermosol (HTS-FRS) and b) University of Wisconsin solution (UW) and a range of preservation temperatures (-4 to -10 oC). We find that there exists an optimum temperature (-4oC) for SCP of rat hepatocytes which yields the highest viability; at this temperature HTS-FRS significantly outperforms UW solution in terms of viability and functional characteristics (secretions and enzymatic activity in suspension and plate culture). With the HTS-FRS solution we show that the cells can be stored for up to a week with high viability (~56%); moreover we also show that the preservation can be performed in large batches (50 million cells) with equal or better viability and no loss of functionality as compared to smaller batches (1.5 million cells) performed in cryovials.

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Section: Discussionmentioning

confidence: 99%

Supercooling as a Viable Non-Freezing Cell Preservation Method of Rat Hepatocytes

et al. 2013

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“…Red blood cells are routinely cryopreserved for blood-banking purposes [10,11]. The process for this results in storage of large blood volumes far beyond what is needed for G6PD testing.…”

Section: Discussionmentioning

confidence: 99%

“…The cryopreserved red blood cells are often used in transfusions and in serological testing. Many blood group reference laboratories have the ability to freeze, reconstitute and analyse red blood cells that were collected months or years earlier [9-11]. Over the years several methods have been used to preserve the metabolic functions of red blood cells [12,13].…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation of glucose-6-phosphate dehydrogenase activity inside red blood cells: developing a specimen repository in support of development and evaluation of glucose-6-phosphate dehydrogenase deficiency tests

Kahn

LaRue

Bansil

et al. 2013

Malar J

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BackgroundGlucose-6-phosphate dehydrogenase (G6PD) deficiency is a common human enzyme deficiency. It is characterized by abnormally low levels of G6PD activity. Individuals with G6PD deficiency are at risk of undergoing acute haemolysis when exposed to 8‒aminoquinoline-based drugs, such as primaquine. For this reason it is imperative to identify individuals with G6PD deficiency prior to administering these anti-malarial drugs. There is a need for the development and evaluation of point-of-care G6PD deficiency screening tests suitable for areas of the developing world where malarial treatments are frequently administered. The development and evaluation of new G6PD tests will be greatly assisted with the availability of specimen repositories.MethodsCryopreservation of erythrocytes was evaluated as a means to preserve G6PD activity. Blood specimens from 31 patients including ten specimens with normal G6PD activity, three with intermediate activity, and 18 with deficient activity were cryopreserved for up to six months.ResultsGood correlation in G6PD activity between fresh and cryopreserved specimens (R2 = 0.95). The cryopreserved specimens show an overall small drop in mean G6PD activity of 0.23 U/g Hb (P=0.23). Cytochemical staining showed that intracellular G6PD activity distribution within the red blood cell populations is preserved during cryopreservation. Furthermore, the mosaic composition of red blood cells in heterozygous women is also preserved for six months or more. The fluorescent spot and the BinaxNOW qualitative tests for G6PD deficiency also showed high concordance in G6PD status determination between cryopreserved specimens and fresh specimens.ConclusionsA methodology for establishing a specimen panel for evaluation of G6PD tests is described. The approach is similar to that used in several malaria research facilities for the cryopreservation of parasites in clinical specimens and axenic cultures. Specimens stored in this manner will aid both the development and evaluation of current and emerging G6PD tests. The availability of G6PD tests is a critical bottleneck to broader access to drugs that confer radical cure of Plasmodium vivax, a requirement for elimination of malaria.

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“…It has been reported by several investigators that membrane permeability is affected by freezing-induced membrane phase transition [10], [40]. Although the addition of cryoprotectant does not prevent liquid-crystal-to-gel membrane phase changes during freezing, the presence of cryoprotectants can decrease the nucleation temperature.…”

Section: Discussionmentioning

confidence: 99%

Slow Freezing Coupled Static Magnetic Field Exposure Enhances Cryopreservative Efficiency—A Study on Human Erythrocytes

et al. 2013

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The aim of this study was to assess the cryoprotective effect of static magnetic fields (SMFs) on human erythrocytes during the slow cooling procedure. Human erythrocytes suspended in 20% glycerol were slowly frozen with a 0.4-T or 0.8-T SMF and then moved to a −80°C freezer for 24 hr. The changes in survival rate, morphology, and metabolites of the thawed erythrocytes were examined. To understand possible cryoprotective mechanisms of SMF, membrane fluidity and dehydration stability of SMF-exposed erythrocytes were tested. For each test, sham-exposed erythrocytes were used as controls. Our results showed that freezing coupled with 0.4-T or 0.8-T SMFs significantly increased the relative survival ratios of the frozen-thawed erythrocytes by 10% and 20% (p<0.001), respectively. The SMFs had no effect on erythrocyte morphology and metabolite levels. However, membrane fluidity of the samples exposed to 0.8-T SMF decreased significantly (p<0.05) in the hydrophobic regions. For the dehydration stability experiments, the samples exposed to 0.8-T SMF exhibited significantly lower (p<0.05) hemolysis. These results demonstrate that a 0.8-T SMF decreases membrane fluidity and enhances erythrocyte membrane stability to resist dehydration damage caused by slow cooling procedures.

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Membrane Stability during Biopreservation of Blood Cells

Cited by 31 publications

References 69 publications

Supercooling as a Viable Non-Freezing Cell Preservation Method of Rat Hepatocytes

Supercooling as a Viable Non-Freezing Cell Preservation Method of Rat Hepatocytes

Cryopreservation of glucose-6-phosphate dehydrogenase activity inside red blood cells: developing a specimen repository in support of development and evaluation of glucose-6-phosphate dehydrogenase deficiency tests

Slow Freezing Coupled Static Magnetic Field Exposure Enhances Cryopreservative Efficiency—A Study on Human Erythrocytes

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