Impact of Ionic Composition of Cryoprotective Medium and Cryopreservation on Human Erythrocyte Sensitivity to Mechanical Stress

Authors

  • Daria I. Aleksandrova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Nina G. Zemlianskykh Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

DOI:

https://doi.org/10.15407/cryo29.04.317

Keywords:

erythrocyte, membrane, mechanical stability, Ca2 , ionic strength, cryopreservation

Abstract

In this research, we have studied the impact of electrolytes and Ca2+ ions on the development of hemolytic damages in human erythrocytes in cryoprotectant solutions under mechanical stress, as well as the effect of freeze-thawing of cells in the presence of glycerol and polyethylene glycol (PEG) on their mechanical stability. The decrease in mechanical stability of cells was found when the salt concentration in extracellular medium increased. At the same time, the absence of electrolytes in cryoprotectant solutions reduced their stability under mechanical stress as well. The Ca2+ introduction into the media increased the cell hemolysis only in the presence of PEG, but not glycerol, apparently due to diff erent effects of these substances on the activity of Ca2+-regulating systems. The introduction of salt of ethylenediaminetetraacetic acid (EDTA) into the media composition reduced the erythrocyte membrane mechanical stability in the presence of the both cryoprotectants, which might be due to the chelator effect on membrane-bound Ca2+. Cryopreservation of erythrocytes increased their sensitivity to mechanical stress, and even the cryoprotectant removal could not restore the cell properties up to the control parameters.

 

Probl Cryobiol Cryomed 2019; 29(4): 317-331

Author Biographies

Daria I. Aleksandrova, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryocytology

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

Department of Cryocytology

References

Balasubramanian SK, Wolkers WF, Bischof JC. Membrane hydration correlates to cellular biophysics during freezing in mammalian cells. Biochim Biophys Acta. 2009; 1788(5): 945-53. CrossRef

Baunbaek M, Bennekou P. Evidence for a random entry of Ca2+ into human red cells. Bioelectrochemistry. 2008; 73(2): 145-50. CrossRef

Bernhardt I, Weiss E. Passive membrane permeability for ions and the membrane potential. In: Bernhardt I, Ellory JC, editors. Red cell membrane transport in health and disease.Berlin, Heidelberg: Springer; 2003. p. 84-109. CrossRef

Ciana A, Achilli C, Balduini C, Minetti G. On the association of lipid rafts to the spectrin skeleton in human erythrocytes. Biochim Biophys Acta. 2011; 1808(1): 183-90. CrossRef

Ciana A, Balduini C, Minetti G. Detergent-resistant membranes in human erythrocytes and their connection to the membrane-skeleton. J Biosci. 2005; 30(3): 317-28. CrossRef

Chasis JA, Mohandas N. Erythrocyte membrane deformability and stability: two distinct membrane properties that are independently regulated by skeletal protein associations. J Cell Biol. 1986; 103(2): 343-50. CrossRef

Esmann M, Fedosova NU, Marsh D. Osmotic stress and viscous retardation of the Na, K-ATPase ion pump. Biophys J. 2008; 94(7): 2767-76. CrossRef

Gulevskyy AK, Riazantsev VV, Kukushkin AI. [Role of the transmembrane potential in impairing the barrier properties of erythrocyte membranes during cryopreservation.] Biull Eksp Biol Med. 1985; 100(12): 690-1. Russian. PubMed

Harano T, Yamaguchi T, Kimoto E. Hemolytic properties of Ca2+-treated human erythrocytes under hydrostatic pressure. J Biochem. 1994; 116(4): 773-7. CrossRef

Ishiguro H, Rubinsky B. Mechanical interactions between ice crystals and red blood cells during directional solidification. Cryobiology. 1994; 31(5): 483-500. CrossRef

Kofanova OA, Zemlyanskikh NG, Ivanova L, Bernhardt I. Changes in the intracellular Ca2+ content in human red blood cells in the presence of glycerol. Bioelectrochemistry. 2008; 73(2): 151-4. CrossRef

Kucherenko YV, Bernhardt I. The study of Ca2+ influx in human erythrocytes in isotonic polyethylene (glycol) 1500 (PEG-1500) and sucrose media. Ukr Biokhim Zh. 2006; 78(6.) 46-52. PubMed

Kummerow D, Hamann J, Browning JA, et al. Variations of intracellular pH in human erythrocytes via K+(Na+)/H+ exchange under low ionic strength conditions. J Membrane Biol. 2000; 176(3): 207-16. CrossRef

Liu F, Mizukami H, Sarnaik S, Ostafin A. Calcium-dependent human erythrocyte cytoskeleton stability analysis through atomic force microscopy. J Struct Biol. 2005; 150(2): 200-10. CrossRef

Logisz CC, Hovis JS. Effect of salt concentration on membrane lysis pressure. Biochim Biophys Acta. 2005; 1717(2), 104-8. CrossRef

Manno S, Takakuwa Y, Mohandas N. Modulation of erythrocyte membrane mechanical function by protein 4.1 phosphorylation. J Biol Chem. 2005; 280(9): 7581-7. CrossRef

Mazur P, Cole KW. Roles of unfrozen fraction, salt concentration, and changes in cell volume in the survival of frozen human erythrocytes. Cryobiology. 1989; (1): 1-29. CrossRef

Moersdorf D, Egee S, Hahn C, Hanf B, Ellory C, Thomas S, Bernhardt I. Transmembrane potential of red blood cells under low ionic strength conditions. Cell Physiol Biochem. 2013; 31(6): 875-82. CrossRef

Mohandas N, Chasis JA. Red blood cell deformability, membrane material properties and shape: regulation by transmembrane, skeletal and cytosolic proteins and lipids. Semin Hematol. 1993; 30(3): 171-92. PubMed

Muldrew K. The salting-in hypothesis of post-hypertonic lysis. Cryobiology. 2008; 57(3): 251-6. CrossRef

Muldrew K, Schachar J, Cheng P, et al. The possible influence of osmotic poration on cell membrane water permeability. Cryobiology. 2009; 58(1): 62-8. CrossRef

Navarro-Prigent MJ, Séguin I, Boivin P, Dhermy D. Study of human erythrocyte membrane protein interactions by selective solubilization of Triton-skeletons. Biol Cell. 1995; 83(1): 33-8. CrossRef

Nunomura W, Takakuwa Y. Regulation of protein 4.1R interactions with membrane proteins by Ca2+ and calmodulin. Front Biosci [Internet]. 2006; [cited 01.03.2018]. (11):1522-39. Available from https://www.bioscience.org/2006/v11/af/1901/fulltext.htm CrossRef

Patton C, Thompson S, Epel D. Some precautions in using chelators to buffer metals in biological solutions. Cell Calcium. 2004; 35(5): 427-31. CrossRef

Pedersen UR, Leidy C, Westh P, Peters GH. The effect of calcium on the properties of charged phospholipid bilayers. Biochim Biophys Acta. 2006; 1758(5); 573-82. CrossRef

Ruan R, Zou L, Sun S, Liu J, Wen L, Gao D, Ding W. Cell blebbing upon addition of cryoprotectants: a self-protection mechanism. PLoS One [Internet]. 2015 [cited 01.03.2018];. 10(4): e0125746. Available from https://journals.plos.org/plosone/article?id=10.1371/journal. pone.0125746 CrossRef

Saragusty J, Gacitua H, Rozenboim I, Arav A. Do physical forces contribute to cryodamage? Biotechnol Bioeng. 2009; 104(4): 719-28. CrossRef

Shpakova NM, Orlova NV, Nipot EE, Aleksandrova DI. [Comparative study of mechanical stress effect on human and animal erythrocytes]. Fiziol Zh. 2015; 61(3): 75-80. Ukrainian. CrossRef

Svetina S, Kuzman D, Waugh RE, Ziherl P, Zeks B. The cooperative role of membrane skeleton and bilayer in the mechanical behaviour of red blood cells. Bioelectrochemistry. 2004; 62(2): 107-13. CrossRef

Takakuwa Y, Ishibashi T, Mohandas N. Regulation of red cell membrane deformability and stability by skeletal protein network. Biorheology. 1990; 27(3-4): 357-65. CrossRef

Takamatsu H, Rubinsky B. Viability of deformed cells. Cryobiology. 1999; 39(3): 243-51. CrossRef

Vácha R, Berkowitz ML, Jungwirth P. Molecular model of a cell plasma membrane with an asymmetric multicomponent composition: water permeation and ion effects. Biophys J. 2009; 96(11): 4493-501. CrossRef

Verstraeten SV, Mackenzie GG, Oteiza PI. The plasma membrane plays a central role in cells response to mechanical stress. Biochim Biophys Acta. 2010; 1798(9): 1739-49. CrossRef

Viallat A, Abkarian M. Red blood cell: from its mechanics to its motion in shear flow. Int J Lab Hematol. 2014; 36(3): 237-43. CrossRef

Zemlianskykh NG. The effects of cryoprotective substances on the mechanical stability and geometric parameters of human erythrocytes. Biophysics. 2018; 63(1): 66-76. CrossRef

Zemlianskykh NG, Babiychuk LA. Cryopreservation in presence of PEG-1500 affects erythrocyte surface characteristics. Probl Cryobiol Cryomed. 2015; 25(2): 104-13. CrossRef

Zemlianskykh NG, Babiychuk LA. The changes in erythrocyte Ca2+-ATPase activity induced by PEG-1500 and low temperatures. Cell and Tissue Biology. 2017, 11(2); 104-10. CrossRef

Zemlyanskikh NG, Khomenko MV. [Human erythrocyte Ca2+-ATPase activity in hypertonic media at low and physiological temperatures]. Biologicheskie Membrany 2006; 23(6): 484-92. Russian.

Zemlyanskikh NG, Kofanova OA. Modulation of human erythrocyte Ca2+-ATPase activity by glycerol: the role of calmodulin. Biochemistry (Mosc). 2006; 71(8): 900-5. CrossRef

Zemlianskykh NG, Kovalenko IF, Babijchuk LA. Peculiarities of modifications in geometric parameters and changes in osmotic fragility of human erythrocytes following their exposure in sucrose and PEG-1500 solutions. Probl Cryobiol Cryomed. 2017; 27(4): 296-310. CrossRef

Downloads

Published

2019-12-17

How to Cite

Aleksandrova, D. I., & Zemlianskykh, N. G. (2019). Impact of Ionic Composition of Cryoprotective Medium and Cryopreservation on Human Erythrocyte Sensitivity to Mechanical Stress. Problems of Cryobiology and Cryomedicine, 29(4), 317–331. https://doi.org/10.15407/cryo29.04.317

Issue

Section

Theoretical and Experimental Cryobiology