Cell-to-Cell Adhesion and Electrical Parameters of Murine Embryo Membranes After Cryopreservation by Vitrification

Authors

DOI:

https://doi.org/10.15407/cryo32.04.256

Keywords:

embryo, adhesion, vitrification, electroporation, electrical breakdown, electrical conductivity, ethylene glycol, cryopreservation

Abstract

In this work, we have studied the impact of various stages of cryopreservation by vitrification in ethylene glycolsucrose medium on plasma membranes adhesive properties of 8-cell murine embryos and their specific electrical conductivity. Embryos were divided into the following experimental groups: the control, group 1 (incubation in vitrification medium) and group 2 (complete cycle of cryopreservation). The embryo exposure to vitrification medium did not affect their ability to cell-to-cell adhesion. After a complete cycle of cryopreservation, no embryo adhesion was observed. Electrical conductivity of embryo membranes was determined using the pulsed conductometry. After incubation in vitrification medium, their resistance to electric pulse was shown to decrease, that was manifested in the phenomenon of irreversible electric breakdown. The average values of electrical conductivity varied within the following ranges: ((12.1 ± 1.5)...(55.5 ± 2.6)), ((28.7 ± 5.7)...(44.± 8.9)), ((31.0 ± 9.3)...(87.9 ± 26.1)) μS/cm in the control, groups 1 and 2, respectively. These findings may be explained by appearance of first structural disorders in lipid bilayer of embryo membranes even at the stage of their incubation in vitrification medium. The lack of adhesive properties of blastomeres after vitrification-warming testified to a damage to the membrane adhesion proteins.

 

Probl Cryobiol Cryomed 2022; 32(4):256–266

Author Biographies

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

Department of Low-Temperature Preservation

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

Department of Low-Temperature Preservation

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

Department of Low-Temperature Preservation

References

Сhehl J. Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research. Acta Physiol Scand. 2003; 177(4): 437-47. CrossRef

Cui XS, Li XY, Shen XH, et al. Transcription profile in mouse four-cell, morula, and blastocyst: Genes implicated in compaction and blastocoel formation. Mol Reprod Dev. 2007; 74(2): 1334-3. CrossRef

De Vries WN, Evsikov AV, Haac BE, et al. Maternal beta-catenin and E-cadherin in mouse development. Development. 2004; 131(18): 4435 45. CrossRef

Emery BR, Miller RL, Carrell DT. Hamster oocyte membrane potential and ion permeability vary with preantral cumulus cell attachment and developmental stage. BMC Dev Biol [Internet]. 2001 10 Oct [Cited 01.11.2021]; 1: 14. Available from: http://www.biomedcentral.com/1471-213X/1/14. CrossRef

Fleming TP, Sheth B, Fesenko I. Cell adhesion in the preimplantation mammalian embryo and its role in trophectoderm differentiation and blastocyst morphogenesis. Front Biosci. 2001; 6(1): 1000–7. CrossRef

Isachenko VV, Grishchenko VI, Ostashko FI, et al. Ultrarapid freezing of embryos of rats and cows without 'classic equilibration'. Problems of Cryobiology. 1994; (3): 3-6.

Kito S, Noguchi Y, Ohta Y, et al. Evaluation of developmental competence of vitrified warmed early cleavage stage embryos and their application for chimeric mouse production. Exp Anim. 2003; 52(2): 179-83. CrossRef

Landel CP. Cryopreservation of mouse gametes and embryos. Guide to Techniques in Mouse Development, Part A: Mice, Embryos, and Cells, 2nd Edition. 2010. 85-105. CrossRef

Mank M. editor. [Developmental biology of mammals. Methods]. Moscow. Мir; 1990. 406 p. Russian.

Mattioli M, Barboni B, Bacci M. Maturation of pig oocytes: Observation on membrane potential. Biol Rreprod. 1990; 43(2): 318-22. CrossRef

Nagy A, Nagy K, Gertsenstein M. Production of mouse chimeras by aggregating pluripotent stem cells with embryos. Guide to Techniques in Mouse Development, Part A: Mice, Embryos, and Cells, 2nd Edition. 2010. 123-49. CrossRef

Nakao K, Nakagata N, Katsuki M. Production of chimeric mice from cryopreserved blastocysts. Exp Anim. 1998; 47(3): 167-71. CrossRef

Shigimaga VA. Pulsed conductometry in a variable electric field: outlook for the development of measurements. Measurement Techniques. 2015; 57(10): 1213-8. CrossRef

Shigimaga VA. Measurements of the capacitance of a biological cell by a pulse method. Measurement Techniques. 2014; 57(2): 213-7. CrossRef

Shigimaga VA. Impulse conductometer for biological cells and liquid media. Measurement Techniques. 2013; 55(11): 1294-300. CrossRef

Smolyaninova YI, Shigimaga VO, Kolesnikova AO, et al. [Determination of electrical conductivity of embryos of mice in preimplantation stages of development after hormonal stimulation of animal ovaries]. Biotechnologia Acta. 2013; 6(1): 105-12. Ukrainian. CrossRef

Smolyaninova YI, Shigimaga VO, Kolesnikova AO. [Influence of hormonal stimulation on morphological and electrical parameters of mouse oocytes]. Animal Biology. 2009; 11(1-2): 329-38. Ukrainian.

Smolyaninova YI, Shigimaga VO, Strikha OA, et al. Effect of cryopreservation stages by vitrification in ethylene glycol and sucrose medium on 2-cell murine embryos electric conductivity. Probl Cryobiol Cryomed. 2013; 23(3): 228-39. Full Text

Smolyaninova YI, Shigimaga VO, Kolesnikova AO, et al. [Influence of the degree of maturity on the electrical conductivity of mouse oocytes]. Animal Biology. 2015; 17(1): 118-25. Ukrainian.

Smolyaninova YI, Shigimaga VO, Kolesnikova AO, et al. Electric conductivity and resistance of mouse oocyte membranes to effect of pulsed electric field in cryoprotectant solutions. Probl Cryobiol Cryomed. 2018; 28 (4): 311-21. CrossRef

Tam PP, Rossant J. Mouse embryonic chimeras: tools for studying mammalian development. Development. 2003; 130(25): 6155-63. CrossRef

Tsong TY. Electroporation of cell membrane. Biophys J. 1991; 60(2): 297-306. CrossRef

Weaver JC. Theory of electroporation: a review. Bioelectrocem Bioenerg. 1996; 41(2): 135-60. CrossRef

White MD, Plachta N. Chapter one - how adhesion forms the early mammalian embryo. Curr Top Dev Biol. 2015; 112: 1-17. CrossRef

Downloads

Published

2023-04-19

How to Cite

Smolyaninova, Y., Shigimaga, V., Popivnenko, L., & Kovalenko, I. (2023). Cell-to-Cell Adhesion and Electrical Parameters of Murine Embryo Membranes After Cryopreservation by Vitrification. Problems of Cryobiology and Cryomedicine, 32(4), 256–266. https://doi.org/10.15407/cryo32.04.256

Issue

Vol. 32 No. 4 (2022): Probl Cryobiol Cryomed

Section

Cryopreservation of Biological Resources

License

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Authors who publish with this journal agree to the following terms:

  • Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
  • Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
  • Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).