Cryopreservation of Equine and Bovine Erythrocytes Using Combined Protective Media

Authors

  • Pavlo Yu. Ulizko Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Olena M Bobrova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Oleg A Nardid 1. Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv 2. V.N. Karazin Kharkiv National University, Kharkiv
  • Pavlo Ðœ. Zubov Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Igor F. Kovalenko Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Vadim M. Kuchkov V.N. Karazin Kharkiv National University, Kharkiv
  • Larisa A. Vodopyanova Kharkiv State Zooveterinary Academy, Kharkiv

DOI:

https://doi.org/10.15407/cryo29.03.255

Keywords:

cryopreservation, equine and bovine red blood cells, combined cryopreservation medium, hemolysis, fluorescent dye

Abstract

A comparative analysis of the effectiveness of equine and bovine erythrocytes’ cryopreservation with the use of one component (20% DMSO) and combined (10% DMSO, 20% PEO-1500) protective medium was carried out. Using fluorescence microscopy and flow cytometry, it has been shown that the 3-DAB fluorescent dye paints the membranes of red blood cells and can be used to evaluate their condition at all cryopreservation stages. It was found that the use of combined protective medium can significantly improve the results of cryopreservation of equine and bovine red blood cells in comparison with one-component media. By cytometric data it has been defined that the parameters of equine erythrocytes after preservation in DMSO-based medium did not correspond the control even after cryoprotectant washing, but after freezing in combined cryoprotective medium and cryoprotectant washing they matched the control. The fluorescence intensification of equine and bovine erythrocytes on fluorescence images and changes in cytograms after mixing cells with 20% DMSO suggests its strong membranotropic influence. Hemolysis level with applying a combined cryoprotective medium of equine and bovine erythrocytes is significantly lower than with the application of 20% DMSO after all cryopreservation stages.

 

Probl Cryobiol Cryomed 2019; 29(3): 255–265.

Author Biographies

Pavlo Yu. Ulizko, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryobiophysics

Olena M Bobrova, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryobiophysics

Oleg A Nardid, 1. Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv 2. V.N. Karazin Kharkiv National University, Kharkiv

Department of Cryobiophysics

Pavlo М. Zubov, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryocytology

Igor F. Kovalenko, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department for Low Temperature Preservation

References

Anchordoguy TJ, Carpenter JF, Crowe JH, et al. Temperature dependent perturbation of phospholipids-bilayers by dimethylsulfoxide. Biochim Biophys Acta. 1992; 1104(1): 117-22. CrossRef

Baskurt OK, Farley RA, Meiselman HJ. Erythrocyte aggregation tendency and cellular properties in horse, human, and rat: a comparative study. American Journal of Physiology-Heart and Circulatory Physiology. 1997; 273(6):H2604-H2612. [Cited 01.07.2019] Available from: https://www.physiology.org/journal/ajpheart CrossRef

Best BP. Cryoprotectant Toxicity: Facts, Issues, and Questions. Rejuvenation Research. 2015; 18(5):422-36. [Cited 01.07.2019] Available from: https://www.liebertpub.com/loi/rej CrossRef

Buriak IA, Zinchenko VD, Dyubko TS, et al. Application of new fluorescent dyes in cryobiological studies. Problems of Cryobiology. 2008; 18(1): 17-21.

Denisova ON, Kuleshova LG, Zemlyanskikh NG, et al. Postthaw morphological changes in animal erythrocytes. Problems of Cryobiology. 2007; 17(2): 150-5.

Denisova ON, Zhegunov GF, Babijchuk LA. Cryopreservation of animal's erythrocytes under dimethyl sulfoxide, polyethylene oxide and glycerol protection. Problems of Cryobiology. 2005; 15(2): 195-201.

Еgorov MI, Dyubko TS, Linnik TP, et al. New fluorescent dyes as fluorescent probes for examination of canine spermatozoa in cryoprotective media. Problems of Cryobiology. 2006. 16(1): 13-23.

Elliott GD, Wang S, Fuller BJ. Cryoprotectants: A review of the actions and applications of cryoprotective solutes that modulate cell recovery from ultra-low temperatures. Cryobiology. 2017; 76:74-91. CrossRef

Holowaychuk MK, Yagi K. Evolution of veterinary transfusion medicine and blood banking. Manual of Veterinary Transfusion Medicine and Blood Banking. 2016 1:1-12. CrossRef

Keeble E, Meredith A, Richardson J, editors. Rabbit medicine and surgery. Veterinary Self-Assessment Color Review Series. Boca Raton: CRC Press, 2016. 243 p. CrossRef

Mair T, Divers T. Equine Internal Medicine. Veterinary selfassessment color review series. Boca Raton: CRC Press; 2015. 399 p. CrossRef

Pogozhykh D, Pakhomova Y, Pervushina O, et al. Exploring the possibility of cryopreservation of feline and canine erythrocytes by rapid freezing with penetrating and non-penetrating cryoprotectants. Stieger K, editor. PLOS ONE. 2017; 12(1): e0169689. [Cited 08.08.2019] Available from: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0169689 CrossRef

Sink C. Practical Transfusion medicine for the small animal practitioner. Jackson: Teton NewMedia, 2004. 110 p.

Ulizko PU, Bobrova OM, Nardid EO, et al. Cryopreservation of animals′ erythrocytes. In: European Research Area: Status, Problems and Prospects. Proceedings of the International Academic Congress; Sept 01–02 2016, Riga, Latvia. Riga; 2016. p. 13–5.

Ulizko PY, Zhegunov GF, Bobrova OM, Zinchenko AV. Influence of mixtures of cryoprotective on safety of cryopreservation of erythrocyte of mammals. Bulletin of problems in biology and medicine. 2011; 3(3): 26-9. Russian.

Won DI, Suh JS. Flow cytometric detection of erythrocyte osmotic fragility. Cytometry Part B: Clin Cytom. 2009;76(2):135-41. CrossRef

Yagi K, Holowaychuk M. Manual of veterinary transfusion medicine and blood banking. Ames: Wiley-Blackwell, 2016. 382 p. CrossRef

Yamamoto A, Saito N, Yamauchi Y, et al. Flow cytometric analysis of red blood cell osmotic fragility. Journal of Laboratory Automation. 2014; 19(5):483-7. [Cited 01.07.2019] Available from: https://journals.sagepub.com/home/jla 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. Problems of Cryobiology and Cryomedicine. 2017; 27(4): 296-310. CrossRef

Zubov PM. Altered lipid asymmetry of membranes of red blood cells of cord and donor blood under Cryopreservation with PEG-1500. Bulletin of problems in biology and medicine. 2013; 2(2): 109-112. Russian.

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Published

2019-09-19

How to Cite

Ulizko, P. Y., Bobrova, O. M., Nardid, O. A., Zubov, P. Ðœ., Kovalenko, I. F., Kuchkov, V. M., & Vodopyanova, L. A. (2019). Cryopreservation of Equine and Bovine Erythrocytes Using Combined Protective Media. Problems of Cryobiology and Cryomedicine, 29(3), 255–265. https://doi.org/10.15407/cryo29.03.255

Issue

Vol. 29 No. 3 (2019): Probl Cryobiol Cryomed

Section

Cryopreservation of Biological Resources

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