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Journal archive > 2008 > N 3 May-June  

INTRACELLULAR FREE CALCIUM CONTENT INCREASE IN THE ERYTHROCYTES TREATED
WITH THE CRYOPROTECTIVE MEDIUM BASED ON POLYETHYLENE GLYCOL 1500 (PEG-1500)

Yu. V. Kucherenko

Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkov;
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Changes in intracellular free calcium content ([Ca2+]i) in human erythrocytes treated with the cryoprotective medium based on low toxic polymer?– polyethylene glycol 1500 (PEG 1500) and then transferred to physiologic salt solution containing 2 mM CaCl2 were studied using fluorescent calcium probe – fura-2. A method of [Ca2+]i calculation with allowance for haemolysis of the cells during the experiment was proposed. It was shown that ignorance of the cell haemolysis resulted­ in significantly higher [Ca2+]i values obtained. Significant time-dependent increase of [Ca2+]i in the cells treated with PEG-1500 cryoprotective medium at +4 °C as well as at +22?°C (without freezing) and then transferred in the 2?mM CaCl2 containing physiological salt solution at +37 °C was observed. Freezing-thawing of the cells treated with the PEG-1500 cryoprotective medium enhanced haemolysis and further accumulation of calcium in the cells. The results of the study prove that the use of PEG-1500-based  cryoprotective medium which does not require washing­ for human erythrocytes will be accompanied by progressive destruction (haemolysis) of the cells in the blood vessels and may have some negative consequences connected with [Ca2+]i increase in the cryopreserved erythrocytes.

Key words: human erythrocytes, calcium, fura-2, cryoprotective medium, polyethylene glycol 1500.

The original article in Russian is available for download in PDF format.

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