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Free Content Mediated Trehalose Un-Loading for Reduced Erythrocyte Osmotic Fragility and Phosphatidylserine Translocation

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Recently, high concentrations of intracellular trehalose (>200mM) were employed to enhance the cryoprotection and desiccation protection of human erythrocytes. However, significant challenges must be overcome if this advancement is to be translated into clinical practice. It is here demonstrated that 247 ± 5 mM intracellular trehalose caused the lysis of 60 ± 2% of erythrocytes upon resuspension in PBS of physiological osmolality (300 mOsm) and caused surviving cells to swell up to 140 ± 2% of isotonic cell volume. Trehalose loaded cells also exhibited 24 ± 1% incidence of phosphatidylserine translocation upon resuspension in 300 mOsm PBS, likely due to loading induced cell swelling. Un-loading of trehalose from erythrocytes using the membrane-permeabilizing biopolymer PP-50 was investigated as a technique to mitigate these damaging effects. After erythrocyte un-loading from 247 ± 5 mM to 39 ± 2 mM intracellular trehalose, cell lysis at 300 mOsm PBS was reduced from 60 ± 2% to 17 ± 3%. Un-loading also reduced cellular incidence of PS translocation in resuspended cells from 24 ± 1% to 13 ± 1%.

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Keywords: BIOPOLYMER; ERYTHROCYTE; OSMOTIC FRAGILITY; PHOSPHATIDYLSERINE; TREHALOSE

Document Type: Research Article

Publication date: September 1, 2011

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  • CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation

    The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.

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