Cryopreservation in Presence of PEG-1500 Affects Erythrocyte Surface Characteristics

Authors

  • Nina G. Zemlianskykh Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine
  • Lyubov A. Babiychuk Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine

DOI:

https://doi.org/10.15407/cryo25.02.104

Keywords:

erythrocytes, membrane, phosphatidylserine, CD44, cryopreservation

Abstract

The studies of changes in surface characteristics of erythrocyte membranes caused by freeze-thawing can be important for assessment of structural and functional integrity of cells and understanding the mechanisms responsible for their damage under extreme conditions. In the research we studied the effect of cryopreservation in presence of exocellular cryoprotectant PEG-1500 on asymmetric distribution of phosphatidylserine in membrane and characteristics of the surface marker CD44 in erythrocyte suspension (the amount of CD44-positive cells and the expression level). It was found that cell amount with the impairments of transmembrane asymmetry of phosphatidylserine increased up to 30% after a short-term exposure of frozen-thawed erythrocytes at 37°C. The decrease in CD44-positive cell amount and lowering of the marker expression were also observed in cryopreserved erythrocyte suspension. Washing of erythrocytes from the cryoprotectant and hemolysis accompanying this process led to restoration of CD44 parameters in suspension of erythrocytes which preserved their integrity. The revealed impairments of surface characteristics of erythrocytes indicate a presence of sublethal membrane injuries in the cells cryopreserved under the protection of PEG-1500. This was true, at least in terms of the changes in the surface marker CD44 indices, since after lysis of cells compromised with sublethal injuries occured during removal of cryoprotectant, the marker indices in erythrocyte suspension did not differ from the control values.


Probl Cryobiol Cryomed 2015; 25(2): 104-113

Author Biographies

Nina G. Zemlianskykh, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine

Department of Cryocytology

Lyubov A. Babiychuk, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine

Department of Cryocytology

References

Belous A.M., Babiychuk L.A., Zemlianskykh N.G. Effect of dosage treatment by polyoxyethyleneoxide mol. m. 1500 to change in shape and permeability of erythrocyte plasma membrane. Doklady of the USSR Academy of Sciences 1989; 7: 59–63.

Borland G., Ross J.A., Guy K. Forms and functions of CD44. Immunology 1998; 93(2): 139–148. CrossRef PubMed

Bratosin D., Estaquier J., Petit F. et al. Programmed cell death in mature erythrocytes: a model for investigating death effector pathways operating in the absence of mitochondria. Cell Death Differ 2001; 8(12):1143–1156. CrossRef PubMed

Daleke D.L. Regulation of phospholipid asymmetry in the erythrocyte membrane. Curr. Opin. Hematol. 2008; 15(3): 191–195. CrossRef PubMed

Devaux P.F., Lopez-Montero I., Bryde S. Proteins involved in lipid translocation in eukaryotic cells. Chem Phys Lipids 2006; 141(1–2): 119–132. CrossRef PubMed

Funaro A., Spagnoli G.C., Momo M. et al. Stimulation of T cells via CD44 requires leukocyte-function-associated antigen interactions and interleukin-2 production. Hum Immunol 1994; 40(4): 267–278. CrossRef

Hale L.P., Singer K.H., Haynes B.F. CD44 antibody against In(Lu)-related p80, lymphocyte-homing receptor molecule inhibits the binding of human erythrocytes to T cells. J. Immunol. 1989; 143(12): 3944–3948. PubMed

Holovati J.L., Wong K.A., Webster J.M., Acker J.P. The effects of cryopreservation on red blood cell microvesiculation, phos-phatidylserine externalization, and CD47 expression. Trans-fusion. 2008; 48(8): 1658–1668. CrossRef PubMed

Iglic A., Veranic P., Jezernik K. et al. Spherocyte shape transformation and release of tubular nanovesicles in human erythrocytes. Bioelectrochemistry 2004; 62(2): 159–161. CrossRef PubMed

Jeremy K.P., Plummer Z.E., Head D.J. et al. 4.1R-deficient human red blood cells have altered phosphatidylserine exposure pathways and are deficient in CD44 and CD47 glycoproteins. Haematologica. 2009; 94(10): 1354–1361. CrossRef PubMed

Lang F., Lang K.S., Wieder T. et al. Cation channels, cell volume and the death of an erythrocyte. Pflugers Arch. 2003; 447(2): 121–125. CrossRef PubMed

Lecak J., Scott K., Young C. et al. Evaluation of red blood cells stored at –80 degrees C in excess of 10 years. Transfusion. 2004; 44(9): 1306–1313. CrossRef PubMed

Lehtonen J.Y., Kinnunen P.K. Poly(ethylene glycol)-induced and temperature-dependent phase separation in fluid binary phospholipid membranes. Biophys. J. 1995; 68(2): 525–535. CrossRef

Lelkens C.C., Noorman F., Koning J.G. et al. Stability after thawing of RBCs frozen with the high- and low-glycerol method. Transfusion. 2003; 43(2): 157–164. CrossRef PubMed

Rudzki Z., Jothy S. CD44 and the adhesion of neoplastic cells. Mol. Pathol. 1997; 50(2): 57–71. CrossRef PubMed

Telen M.J. Erythrocyte blood group antigens: not so simple after all. Blood. 1995; 85(2): 299–306. PubMed

Telen M.J. Red blood cell surface adhesion molecules: their possible roles in normal human physiology and disease. Semin. Hematol. 2000; 37(2): 130–142. CrossRef

Xu Z., Duffett L., Tokessy M. et al. Anti- AnWj causing acute hemolytic transfusion reactions in a patient with aplastic ane-mia. Transfusion. 2012; 52(7): 1476–1481. CrossRef PubMed

Zemlianskikh N.G. About the problem of erythrocyte stability, cryopreserved under polyethyleneglycol protection. Bulletin of the problems of biology and medicine. 2007; 3: 32–37.

Zemlyanskikh N.G., Khomenko M.V. Human erythrocyte Ca2+-ATPase activity in hypertonic media at low and physiological temperatures. Biol Membrany 2006; 23(5): 375–383.

Zemlyanskikh N.G., Kofanova O.A. Modulation of human erythrocyte Ca2+-ATPase activity by glycerol: the role of calmodulin. Biochemistry (Moscow). 2006; 71(8): 900–905. CrossRef

Downloads

Published

2015-06-20

How to Cite

Zemlianskykh, N. G., & Babiychuk, L. A. (2015). Cryopreservation in Presence of PEG-1500 Affects Erythrocyte Surface Characteristics. Problems of Cryobiology and Cryomedicine, 25(2), 104–113. https://doi.org/10.15407/cryo25.02.104

Issue

Section

Theoretical and Experimental Cryobiology