Bìol. Tvarin, 2017, Volume 19, Issue 2, pp. 64–69


A.Mazur, E. Ye. Nipot, N. V. Orlova, N. M. Shpakova

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Institute for Problems of Cryobiology and Cryomedicine NAS of Ukraine,
23 Pereyaslavska str., Kharkiv 61016, Ukraine, This email address is being protected from spambots. You need JavaScript enabled to view it.

The sensitivity of mammalian erythrocytes to post-hypertonic shock which is one of the main factors of an injury of the cells during their cryopreservation was studied in this work. The research aim was to comparatively investigate the sensitivity of bovine and ram erythrocytes to the action of post-hypertonic shock with varying NaCl concentration in the dehydration medium and temperature.

Post-hypertonic shock was initiated by transferring the erythrocytes from hypertonic media (1.0–2.5 mol/L NaCl) to an isotonic solution (0.15 mol/L NaCl) at different temperatures (0 to 37 °C). The level of hemolysis was determined spectrophotometrically at a wavelength of 543 nm.

It has been shown that with a rise of salt concentration in the dehydration medium, the level of post-hypertonic hemolysis of bovine and ram erythrocytes increased at the both temperatures 37 and 0 °C. It should be noted that the ram erythrocytes were more sensitive to the action of post-hypertonic shock at 37 °C and the bovine ones were more sensitive at 0 °C.

The analysis of the temperature dependences of post-hypertonic hemolysis of bovine and ram erythrocytes showed a significant difference for the cells of these mammals. Thus, the temperature dependence of ram erythrocytes had a minimum at 20 °C, and bovine ones exhibited the same level of damage within the temperature range of 0–10 °C and the growth of hemolysis with a further increase in temperature. It should be noted that at the temperatures above 15 °C the post-hypertonic hemolysis of ram erythrocytes was almost 2 times higher than the level of damage for bovine cells.

The revealed features of the sensitivity of bovine and ram erythrocytes to the action of post-hypertonic shock could be explained by the differences in the mechano-elastic properties of their membranes, which might be due to the peculiarities of the formation of membrane clusters and their interactions within the cytoskeleton-membrane complex.


  1. Alexandrova D. I., Orlova N. V., Shpakova N. M. A comparative study of the sensitivity of previously dehydrated human and bovine red blood cells to hypertonic. Problems of cryobiology, 2007, no. 4, pp. 327–334. (in Russian)
  2. Al-Qarawi A. A., Mousa H. M. Lipid concentrations in erythrocyte membranes in normal, starved, dehyrated and rehydratedcamels (Camelus dromedarius), and in normal sheep (Ovis aries) and goats (Capra hircus). Eur. J. Arid Environ., 2004, vol. 59, no. 4, pp. 675–683. https://doi.org/10.1016/j.jaridenv.2004.02.004
  3. Belous A. M., Bondarenko V. A., Babiychyk L. A. Single mechanism of cell damage in thermal shock, freezing and posthypertonic lysis. Cryobiology, 1985, no. 2, pp. 25–32. (in Russian)
  4. Benga G. Comparative studies of water permeability of red blood cells from humans and over 30 animal species: an overview of 20 years of collaboration with Philip Kuchel. Eur. Biophys. J., 2013, vol. 42, no. 1, pp. 33–46. https://doi.org/10.1007/s00249-012-0868-7
  5. Ding J., Niu S., Chen Z., Zhang T., Griffith B. P., Wu Z. J. Shear-induced hemolysis: species differences. Artif Organs., 2015, vol. 39, no. 9, pp. 795–802. https://doi.org/10.1111/aor.12459
  6. Florin-Christensen J., Suarez C. E., Florin-Christensen M., Wainszelbaum M., Brown W. C., McElwain T. F., Palmer G. H. An unique phospholipid organization in bovine erythrocyte membranes. Proc. Natl. Acad. Sci. USA, 2001, vol. 98, no. 14, pp. 7736–7741. https://doi.org/10.1073/pnas.131580998
  7. Goltsev A. N., Gordienko E. A., Babiychuk G. A., eds. Current problems of cryobiology and cryomedicine. Kharkov, 2012, Rayder Publ., 2012, 767 p. (in Russian)
  8. Gordienko E. A., Tovstyak V. V. Physics of biomembranes. Kyiv, Naukova dumka, 2009, 277 p. (in Ukrainian)
  9. Lang F., Foler M. Erythrocytes: Physiology and Pathophysiology. London, Imperial College Press Publ., 2012, 443 p. https://doi.org/10.1142/p740
  10. Matei H., Frentescu L., Benga Gh. Comparative studies of the protein composition of red blood cell membranes from eight mammalian species. J. Cell. Mol. Med., 2000, vol. 4, no. 4, pp. 270–276. https://doi.org/10.1111/j.1582-4934.2000.tb00126.x
  11. Muldrew K. The salting-in hypothesis of post-hypertonic lysis. Cryobiology, 2008, vol. 57, no. 3, pp. 251–256. https://doi.org/10.1016/j.cryobiol.2008.09.007
  12. Nezar A., Melizi M., Belabbas H. Species determination using the red blood cells morphometry in domestic animals. Vet. World, 2016, vol. 9, no. 9, pp. 960–963. https://doi.org/10.14202/vetworld.2016.960-963
  13. Noha A. S. Salidroside as a novel protective agent to improve red blood cell cryopreservation. PLoS One, 2016, vol. 11, no. 9, pp. 1–12. https://doi.org/10.1371/journal.pone.0162748
  14. Patelaros S. V., Synchikova O. P. Osmotic behavior of erythrocytes in post-hypertensive lysis. Problems of cryobiology, 1994, no. 3, pp. 35–40. (in Russian)
  15. Semyonova E. A., Ershova N. A., Ershov S. S., Peculiarities of postgypertensive lysis of erythrocytes of some mammals. Problems of cryobiology and cryomedicine, 2016, vol. 26, no. 1, pp. 73–83. (in Russian)
  16. Shpakova N. M., Ershov S. S. Hypertonic cryohemolysis of mammalian erythrocytes. Problems of cryobiology, 2006, no. 3, pp. 286–291. (in Russian)
  17. Shpakova N. M. The temperature and osmotic resistance of red blood cells of different species of mammals. Dr. biological sci. diss. Kharkiv, 2014, 392 p. (in Russian)

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