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ISSN 2522-9028 (Print)
ISSN 2522-9036 (Online)
DOI: https://doi.org/10.15407/fz

Fiziologichnyi Zhurnal

is a scientific journal issued by the

Bogomoletz Institute of Physiology
National Academy of Sciences of Ukraine

Editor-in-chief: V.F. Sagach

The journal was founded in 1955 as
1955 – 1977 "Fiziolohichnyi zhurnal" (ISSN 0015 – 3311)
1978 – 1993 "Fiziologicheskii zhurnal" (ISSN 0201 – 8489)
1994 – 2016 "Fiziolohichnyi zhurnal" (ISSN 0201 – 8489)
2017 – "Fiziolohichnyi zhurnal" (ISSN 2522-9028)

Fiziol. Zh. 2014; 60(6): 46-53


PROPERTIES OF ERYTHROCYTES WITH LOW DEGREE OF INTEGRITY AFTER FREEZING WITH NON-PENETRATING AND PENETRATING CRYOPROTECTANTS

V.V. Ramazanov, E.L. Volovelskaya, V.A. Koptelov, V.A. Bondarenko

    Institute for Problems of Cryobiology and CryomedicineNational Academy of Sciences of Ukraine, Kharkov, Ukraine
DOI: https://doi.org/10.15407/fz60.06.046


Abstract

Osmotic, antioxidant and morphological characteristics of erythrocytes frozen in liquid nitrogen (-196°C) in the medium containing sucrose, dextran and DMSO were studied. During freezing in the medium containing sucrose and sucrose and dextran there is noted the disorder in osmotic and morphological characteristics of the rest of cells after washing-out from cryopreservative. We have not revealed a significant change in concentration of malondialdehyde and the indices of activity of glutathione-dependent enzymes (glutathione reductase and glutathione peroxidase). Inclusion of DMSO into the mentioned media results in the decreased degree of erythrocytes damage during freezing and preservation of osmotic, antioxidant and morphological indices of the rest of cells independent of the degree of their integrity after washing-out from cryopreservative. The obtained results indicate that in order to preserve the properties of erythrocytes during freezing the cryopreservative should include the combination of non-penetrating and penetrating cryoprotectants.

Keywords: erythrocytes, osmotic, antioxidant and morphologicalproperties, freezing, combined cryopreservative.

References

  1. Pellerin-Mendes C., Million L., Marchand-Arvier M., et al. In vitro study of the protective effect of trehalose and dextran during freezing of human red blood cells in liquidnitrogen. Cryobiology. 1997; 35(2): 173–86. CrossRef PubMed
  2.  
  3. Sputtek A., Singbartl G., Langer R., et al. Cryopreservation of erythrocytes using hydroxyethyl starch: in vivo results of an autologous retransfusion model in humans. Beitr Infusionsther Transfusionsmed. 1994; 32: 44–7. PubMed
  4.  
  5. Wagner C.T., Martowicz M.L., Livesey S.A., Connor J. Biochemical stabilization enhances red blood cell recovery and stability following cryopreservation. Cryobiology. 2002; 45(2): 153–66. CrossRef  
  6. Ramazanov V.V., Dejneko T.I., Volovelskaya E.L., et al. Properties of erythrocytes frozen in combined medium with polyethylene glycol and dimethylsulfoxide. Biotechnology. 2012; 5(2): 106–14 [Ukrainian].
  7.  
  8. Bakhach J. The cryopreservation of composite tissues. Principles and resent advancement on cryopreservation of different type of tissues. Organogenesis. 2009; 5(3): 119–26. CrossRef PubMed PubMedCentral
  9.  
  10. Mazur P., Cole K.W. Roles of unfrozen fraction, salt concentration, and changes in cell volume in the survival of frozen human erythrocytes. Cryobiology. 1989; 26(1): 1–29. CrossRef  
  11. Pegg D.E. The effect of cell concentration on the recovery of human erythrocytes after freezing and thawing in the presence of glycerol. Cryobiology. 1981; 18(3): 221–8. CrossRef  
  12. Ramazanov V.V. Effect of combined media on damage of erythrocytes frozen with different hematocrit values. Problems of Cryobiology. 2006; 16 (2): 155–163 [Ukrainian].
  13.  
  14. Romano L., Passow H. Characterization of anion transport system in trout red blood cell. Am. J. Physiol. 1984; 246(3): 330–8. CrossRef PubMed
  15.  
  16. Ramazanov V.V., Zabrodskiy R.F., Nayduk Ya.Yu., Bondarenko V.A. H+ ion transport system functioning upon erythrocyte membrane modifications under conditions modeling freezing conditions. Visnyk Problem Biologii iMedytsyny. 2010; Iss 3: 186–192 [Ukrainian].
  17.  
  18. Wieth J.O., Bjerrum P.J., Borders C.L. Irreversible inactivation of red cell chloride exchange with phenylglyoxal, and arginine– specific reagent. J Gen. Physiol. 1982; 79(2): 283–312. CrossRef PubMed
  19.  
  20. Beutler E. Red cell metabolism. A manual of biochemical methods. New York: Grune&Stratton; 1975. 160 p. PubMedCentral
  21.  
  22. Vladimirov Yu.A., Archakov A.I. Lipid peroxidation and biological membranes. Moscow: Nauka; 1972. 270 p.[Russian].
  23.  
  24. Razygraev A.V., Arutunyan A.V. Determination of glutathione peroxidase activity in human blood serum using hydrogen peroxide and 5,5'-dithiobis (nitrobenzoic acid). Klin Lab Diagnostika. 2006; (6): 13–6 [Russian].
  25.  
  26. Yusupova L.B. On increasing the accuracy of determination of glutathione reductase activity in erythrocytes. Lab Delo. 1989; (4): 19–21 [Russian].
  27.  
  28. Ashmarin I.P., Vasiliev I.P., Ambrosov V.A. Quick methods for statistical processing and experiment designing. Leningrad; 1975. 76 p. [Russian].
  29.  
  30. Ramazanov V.V., Bondarenko V.A. Osmotic properties of erythrocytes frozen in media containing non-penetrating and penetrating cryoprotectants. Problems of Cryobiology. 2010; 20(1): 47–58 [Ukrainian].
  31.  
  32. Bartosz G., Niewiarowska J., Judkiewicz L. Decreased deformability in aging erythrocytes. Biochim Biophys Acta. 1982; 693(1): 262–4. CrossRef  
  33. Imanishi H, Nakai T, Abe T, Takino T. Glutathione-linked enzyme activities in red cell aging. Clin Chim Acta. 1986;159(1): 73–6. CrossRef  

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