Effect of Cryopreservation Stages by Vitrification in Ethylene Glycol and Sucrose Medium on 2-Cell Murine Embryos Electric Conductivity
Keywords:mouse embryo, electroporation, electric breakdown, vitrification, ethylene glycol, cryopreservation
In this study the values of specific electric conductivity of 2-cell murine embryos after their exposure in ethylene glycol and sucrose medium and complete cryopreservation cycle by vitrification were determined using the method of impulse conductometry. It has been shown that embryo exposure in vitrifying medium led to an increase in their electric conductivity values in comparison with the control ones ((2.24 Â± 0.64) - (3.86 Â± 0.68)x10â€“3 S/m and (1.54 Â± 0.110 - 96.12 Â± 0.34)x10â€“3 S/m; respectively). Extension of exposure time from 1.5 min to 3 min led to a decrease in embryo plasma membrane resistance to pulse electric field action. The investigation showed that electric conductivity could be a sensitive diagnostic cell parameter reflecting disturbances in cell morphofunctional integrity at various stages of cryopreservation. Probl Cryobiol Cryomed 2013; 23(3):228â€“239.
Developmental biology of mammals. Methods / Ed. by M. Monk. Academic Press, 1990.
Isachenko V.V., Grishchenko V.I., Ostashko F.I. et al. Ultrarapid freezing of embryos of rats and cows without â€˜classic equilibrationâ€™. Problems of Cryobiology 1994; 3: 3â€“6.
Kolesnikova A.A., Shigimaga V.A., Smolyaninova E.I. Effect of the stimulated maturation and fertilization on the electric conductivity and fertilization of mouse oocytes. Fundamentalnye Issledovaniya 2013; (Part 4, N4): 896â€“899.
Krivoharchenko A.S., Vilyanovich L.I., Serobyan G.A. et al. Survival of mouse embryos after ultra-rapid freezing in different ways using different cryoprotectants. Problemy Reproduktsii 1995; 4: 13â€“18.
Pogorelov A.G., Goldstein D.V., Smolyaninova E.I., Sakharova N.Yu. Electron probe microanalysis of the intracellular concentration of potassium in the cells of the early mouse embryo in the preimplantation period. Doklady Akademii Nauk. â€“ 2005; 400 (2): 276â€“278.
SmolyanÑ–nova E.I., Strikha O.A., Shigimaga V.O. et al. Determination of electric conductivity of murine embryos of preâ€“implantation streges after hormonal stimulation of animal oviducts. Biotechnologia Acta. â€“ 2013 ; 6, N1: 105â€“112.
SmolyanÑ–nova E.I., Shigimaga V.O., KolesnÑ–kova A.O. Effect of hormonal stimulation on morphofunctional and electric parameters of murine oocytes. BÑ–ologÑ–ya Tvarin 2009; 11 (1â€“2): 329â€“338 .
Smolyaninova E.I., Pishko O.V., Lisina E.G. et al. Analysis of the effect of different stages of cryopreservation by vitrification in ethylene glycol â€“ sucrose medium on the viability of twoâ€“cell mouse embryos. Problems of Cryobiology 2007; 17 (4): 385â€“393.
Smolyaninova E.I., Pogorelov A.G., Lisina E.G. et al. Effect of freezing medium on viability and the cationic composition of early mouse embryos. Problems of Cryobiology 2005; 15(3): 3â€“10.
Smolyaninova E.I., Khromenkova O.B., Zhernoklev G.V., Pischko O.V. Effect of equilibration in the freezing medium on osmotic stability and viability of 8-cell mouse embryos. Problems of Cryobiology 2004; 1: 3â€“11.
Smolyaninova E.I., Khromenkova O.B., Zhernoklev G.V. Effect of different stages of cryopreservation method by ultra-rapid freezing on osmotic stability and morphofunctional integrity of mouse embryos. Problems of Cryobiology 2001; 2: 49â€“55.
Chizmadzhiev Yu.A., Chernomordik L.V., Pastushenko V.F., Abidor I.G. Electric breakdown of bilayer lipid membranes. Itogi Nauki i Tekhniki. Series: Biophysics of Membranes 1982; 2. â€“ 308 p.
Shigimaga V.A., Megel Yu.E. Method for determining the conductivity of oocytes and embryos in different conditions of the dielectric environment. In: New solutions in modern technology. Collection of papers. â€“ Kharkov, 2011; 9: 140â€“144.
Shigimaga V.A. Determination of conductivity of embryonic cells of animals. Problemy Bioniki 2003; (59): 60â€“64.
Patent Nr 34333 of Ukraine, IPC G01N33/483. Method of determination of resistance of single cell membrane in cryoprotectant solution. V.O. Shigimaga, E.I. SmolyanÑ–nova, Applied by and owned by Ð†PC&C of the National Academy of Sciences of Ukraine. Nr.U200802374; Filed 25.02.2008; Published 11.08.2008, Bulletin Nr. 15.
Ð¡hehl J. Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research. Acta Physiol Scand 2003; 177, (4): 437â€“447.
Cobo A., de los Santos M.J., Castello D. et al. Outcomes of vitrified early stages and blastocyst-stage embryos in cryopreservation program: evaluation of 3,150 warming cycles. Fertil Steril 2012; 98 (5): 1138â€“1146.
Day M.L., Johnson M.H., Cook D.J. A cytoplasmic cell cycle controls the activity of a K+ channel in preimplantation mouse embryos. EMBO J 1998; 17 (7): 1952â€“1960.
Dovey S. Oocyte cryopreservation: advances and draw back. Minerva Ginecol 2012; 64 (6): 485â€“500.
Emery B.R., Miller R.L., Carrell D.T. Hamster oocyte membrane potential and ion permeability vary with prenatal cumulus cell attachment and development and developmental stage. BMC Dev Biol 2001; 1: 14.
Gosden R. Cryopreservation: a cold look at technology for fertility preservation. Fertil Steril 2013; 96 (2): 264â€“268.
Guide to electroporation and electrofusion / Ed. by: D.C. Chang, B.M. Chassy, J.A. Saunders, A.E. Sowers. London: Academic Press, 1992.
Harper J., Magli M.C., Lundin K. et al. When and how should new technology be introduced into the IVF laboratory. Hum Reprod 2012; 27 (2): 303â€“313.
Jin B., Mochida K., Ogura A. et al. Equilibrium vitrification of mouse embryos at various developmental stages. Mol Reprod Dev 2012; 79 (11): 785â€“794.
Lin J., Phy J., Yeomans E. Theoretical considerations regarding slow cooling and vitrification. Theriogenology 2012; 78 (8): 1641â€“1652.
Mukaida T., Oka C. Vitrification of oocytes, embryos and blastocysts. Best Pract Res Clin Obst Gynaecol 2012; 26 (6): 789â€“803.
Pogorelov A.G., Katkov I.I., Smolyaninova E.I. Goldshtein D.V. Changes in intracellular potassium and sodium content of 2-cell mouse embryos induced by exposition to vitrification concentrations of ethylene glycol. CryoLetters 2006; 27 (2): 87â€“98.
Rols M. P. Mechanism by which electroporation mediates DNA migration and entry into cells and targeted tissues. Methods Mol Biol 2008; 423 (1): 19â€“33.
Rols M.P. Electropermeabilization, a physical method for the delivery of therapeutic molecules into cells. Biochim Biophys Acta 2006; 1758 (3): 423â€“428.
Shigimaga V.A. Impulse conductometer for biological cells and liquid media. Measurement Techniques 2013; 55 (11): 1294â€“1300.
Tsong T.Y. Electroporation of cell membrane. Biophys J 1991; 60 (2): 297â€“306.
Trimarchi J.R., Lui L., Smith P.J.S., Keefe D.L. Noninvasive measurement of potassium efflux as an early indicator of cell death in mouse embryos. Biol Reprod 2000; 63 (3): 851â€“857.
Weaver J.C. Theory of electroporation: a review. Bioelectrochem Bioenerg 1996; 41 (2): 135â€“160.
Zimmermann U. Electric field mediated fusion and related electrical phenomena. Biochim Biophys Acta 1982; 694 (3): 227â€“277.
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Copyright (c) 2020 Evgeniya I. Smolyaninova, Viktor A. Shigimaga, Oksana A. Strikha, Lyudmila I. Popivnenko, Ekaterina G. Lisina
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