Українська English

ISSN 2522-9028 (Print)
ISSN 2522-9036 (Online)

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. 2024; 70(2): 59-66


O.E. Nipot, N.A. Yershova, O.O. Chabanenko, P.M. Zubov, N.M. Shpakova

    Institute of Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine


The search for protective substances that can be used during red blood cell thawing and the study of their effects on red blood cells contribute to increasing the number and quality of viable cells after the cryopreservation cycle. We studied the effect of posthypertonic shock and amphiphilic compounds on the shape and eryptosis of human erythrocytes. The method of flow cytometry was used, this allows analyzing two parameters simultaneously, which increases the efficiency of research. The shape was assessed by the sphericity index (SphI), and eryptosis by the redistribution of phosphatidylserine to the membrane outer surface. It has been shown that sodium decylsulfate and chlorpromazine reduce erythrocyte damage in posthypertonic shock by 3.6 and 4.2 times, respectively. Sodium decylsulfate helps to preserve the shape of cells (SphI coefficient remains the same), while when chlorpromazine is used, the shape changes towards spherical (SphI coefficient changes 2 times). The study of the level of Annexin V FITC binding to phosphatidylserine in outer layer of membrane revealed a concentration-dependent increase in fluorescence when sodium decylsulfate was used, indicating a disorder of the bilayer asymmetry. In contrast, chlorpromazine did not change the distribution of phosphatidylserine. Comparison of two parameters of cell viability - the sphericity coefficient and anexin binding - allowed us to choose the conditions that are optimal for the use of the studied protective substances. Namely, it is advisable to use the lowest effective concentration of sodium decylsulfate (200 mcmol/l) for protective purposes. This ensures the preservation of the cell shape and minimal impact on the membrane asymmetry.

Keywords: erythrocyte; posthypertonic shock; amphiphilic compounds; sphericity coefficient; phosphatidylserine; flow cytometry.


  1. Lovelock JE. The haemolysis of human red blood-cells by freezing and thawing. Biochim Biophys Acta. 1953;10:414-26. CrossRef PubMed
  2. Klbik I. On the involvement of cell volume regulation mechanisms in post-hypertonic lysis and slow-freezing injury of human erythrocytes and its broader cryobiological significance. BioRxiv. 2023 Apr 28:29. CrossRef PubMed
  3. Muldrew K. The salting-in hypothesis of post-hypertonic lysis. Cryobiology. 2008;57(3):251-56. CrossRef PubMed
  4. Chabanenko O, Yershova N, Shpakova N. Adequacy of posthypertonic shock model to real cryopreservation conditions during deglycerolization of erythrocytes. Proceedings of the 57th annual meeting of the Society for Cryobiology «CRYO-2020». 21-23 July 2020, USA. Cryobiology. 2020; 97: 276. CrossRef
  5. Orlova NV, Shpakova NM Mechanism of protective effect of amphiphilic compounds during hypertonic hemolysis of erythrocytes. Fiziol Zh. 2006;52(5):55-61. [Ukrainian].
  6. Yershova NA, Chabanenko OO, Shpakova NМ, Nipot OЕ, Orlova NV. Effect of trifluoroperazine and sodium decyl sulfate on the posthypertensive shock of human and rabbit red blood cells. Fiziol Zh. 2022;68(1):62-8. [Ukrainian]. CrossRef
  7. Alvesa I, Stanevab G, Tessierac C, Salgadod GF, Nussac P. The interaction of antipsychotic drugs with lipids and subsequent lipid reorganization investigated using biophysical methods. Biochim Biophys Acta - Biomembran. 2011;1808(8):2009-18. CrossRef PubMed
  8. Habibi S, Lee HY, Moncada-Hernandez H, Gooding J, Minericka AR. Impacts of low concentration surfactant on red blood cell dielectrophoretic responses. Biomicrofluidics. 2019;13(5):054101. CrossRef PubMed PubMedCentral
  9. Braasch D. The relation between erythrocyte deformability, cell shape, and membrane surface tension. Pflüg Arch. 1969; 313:316-20. CrossRef PubMed
  10. Geekiyanage N, Sauret E, Saha S, Flower R, Gu Y. Modelling of red blood cell morphological and deformability changes during in-vitro storage. Applied Sciences. 2020; 10(9):3209. CrossRef
  11. Ebrahimi S, Bagchi P. A computational study of red blood cell deformability effect on hemodynamic alteration in capillary vessel networks. Sci Rep. 2022;12; 4304. CrossRef PubMed PubMedCentral
  12. Bevers EM, Williamson PL. Getting to the outer leaflet: physiology of phosphatidylserine exposure at the plasma membrane. Physiol Rev. 2016; 96: 605-45. CrossRef PubMed
  13. Alghareeb SA, Alfhili MA, Fatima S. Molecular Mechanisms and pathophysiological significance of eryptosis. Int J Mol Sci. 2023; 24(6):5079. CrossRef PubMed PubMedCentral
  14. Hagerstrand H, Holmström TH, Bobrowska-Hägerstrand M, Eriksson JE, Isomaa B. Amphiphile-induced phosphatidylserine exposure in human erythrocytes. Mol Membran Biol. 1998; 15(2):89-95. CrossRef PubMed
  15. Piagnerelli M, Zouaoui Boudjeltia K, Brohee D, Vereerstraeten A, Piro P, Vincent JL, Vanhaeverbeek M. Assessment of erythrocyte shape by flow cytometry techniques. J Clin Pathol. 2007; 60(5):549-54. CrossRef PubMed PubMedCentral
  16. Freikman I, Amer J, Ringel I, Fibach E. A flow cytometry approach for quantitative analysis of cellular phosphatidylserine distribution and shedding. Anal Biochem. 2009;393(1):111-6. CrossRef PubMed
  17. Shpakova NM, Semionova EA, Kovalenko IF, Iershova NA, Orlova NV. Morphological peculiarities of temperature and osmotic response of erythrocytes in presence of chloropromazine. Fiziol Zh. 2017;63(5):62-9. [Ukrainian]. CrossRef
  18. Ficarra S, Russo A, Barreca D, Giunta E, Galtieri A, Tellone E Short-term effects of chlorpromazine on oxidative stress in erythrocyte functionality: activation of metabolism and membrane perturbation. Oxid Med Cell Longev. 2016;2016:2394130. CrossRef PubMed PubMedCentral
  19. Gienger J, Gross H, Ost V, Bär M, Neukammer J. Assessment of deformation of human red blood cells in flow cytometry: measurement and simulation of bimodal forward scatter distributions. Biomed Opt Express. 2019;10(9):4531-50. CrossRef PubMed PubMedCentral
  20. Ghosh S, Chakraborty I, Chakraborty M, Mukhopadhyay A, Mishra R, Sarkar D. Evaluating the morphology of erythrocyte population: An approach based on atomic force microscopy and flow cytometry. Biochim Biophys Acta. 2016;1858(4):671-81. CrossRef PubMed
  21. Huisjes R, Bogdanova A, Solinge WW, Schiffelers RM, Kaestner L, Wijk R. Squeezing for life - properties of red blood cell deformability. Front Physiol. 2018; 9: 656. CrossRef PubMed PubMedCentral
  22. Lundbæk JA. Lipid bilayer-mediated regulation of ion channel function by amphiphilic drugs. J Gen Physiol. 2008;131(5):421-29. CrossRef PubMed PubMedCentral
  23. Wesseling MC, Wagner-Britz L, Huppert H, Hanf B, Hertz L, Nguyen DB, Bernhardt I. Phosphatidylserine exposure in human red blood cells depending on cell age. Cell Physiol Biochem. 2016;38(4):1376-90. CrossRef PubMed

© National Academy of Sciences of Ukraine, Bogomoletz Institute of Physiology, 2014-2024.