Erythrocyte swelling after rapid dilution of cryoprotectants and its prevention.
These in vitro studies on canine red blood cells confirm that cell swelling occurs after rapid dilution of Me2SO and glycerol. Cells loaded with a penetrating cryoprotectant in a medium with low Na+, high K+ composition present significantly less swelling after rapid dilution of the cryoprotectant than cells exposed to an electrolyte medium characterized by high Na+, low K+ composition. The osmotic cell stress during rapid dilution of Me2SO can be completely counteracted by the simultaneous use of the nonpenetrating sorbitol during exposure and loading. However, the addition of sorbitol is of no important benefit when glycerol is used as the intracellular cryoprotectant. This is probably due to the slower elution of glycerol. Thus utilizing a perfusion solution containing sorbitol during loading and dilution of Me2SO reduces the osmotic injury and may greatly improve the survival prospects of cryopreserved organs by avoiding "out-flow" block.
D000818:Animals; D001793:Blood Preservation / Q000379:methods*; D002021:Buffers; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004285:Dogs; D004905:Erythrocyte Aging / Q000187:drug effects*; D004909:Erythrocyte Indices / Q000187:drug effects; D004912:Erythrocytes / Q000502:physiology*; D005260:Female; D005990:Glycerol / Q000494:pharmacology; D007473:Ion Channels / Q000187:drug effects; D008954:Models, Biological; D009996:Osmotic Fragility / Q000187:drug effects; D011188:Potassium / Q000378:metabolism / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism / Q000494:pharmacology; D013012:Sorbitol / Q000494:pharmacology
D002021:Buffers; D003451:Cryoprotective Agents; D007473:Ion Channels; D013012:Sorbitol; D012964:Sodium; D005990:Glycerol; D011188:Potassium; D004121:Dimethyl Sulfoxide