Loading equine oocytes with cryoprotective agents captured with a finite element method model.
Abstract: Cryopreservation can be used to store equine oocytes for extended periods so that they can be used in artificial reproduction technologies at a desired time point. It requires use of cryoprotective agents (CPAs) to protect the oocytes against freezing injury. The intracellular introduction of CPAs, however, may cause irreversible osmotic damage. The response of cells exposed to CPA solutions is governed by the permeability of the cellular membrane towards water and the CPAs. In this study, a mathematical mass transport model describing the permeation of water and CPAs across an oocyte membrane was used to simulate oocyte volume responses and concomitant intracellular CPA concentrations during the exposure of oocytes to CPA solutions. The results of the analytical simulations were subsequently used to develop a phenomenological finite element method (FEM) continuum model to capture the response of oocytes exposed to CPA solutions with spatial information. FEM simulations were used to depict spatial differences in CPA concentration during CPA permeation, namely at locations near the membrane surface and towards the middle of the cell, and to capture corresponding changes in deformation and hydrostatic pressure. FEM simulations of the multiple processes occurring during CPA loading of oocytes are a valuable tool to increase our understanding of the mechanisms underlying cryopreservation outcome.
© 2021. The Author(s).
Publication Date: 2021-10-06 PubMed ID: 34615933PubMed Central: PMC8494918DOI: 10.1038/s41598-021-99287-9Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research studies a mathematical model to understand the process of protecting horse eggs (oocytes) from freezing damage for long-term storage using cryoprotective agents (CPAs). The understanding and modelling of this process of cryopreservation are critical for the optimisation of artificial reproduction technologies.
Background
- The study focuses on the process of cryopreservation, a method of preserving biological tissue by cooling it to very low temperatures, and the use of cryoprotective agents (CPAs) that protect the cells from freezing damage.
- Though effective, the introduction of CPAs into cells can cause irreversible osmotic damage, necessitating proper understanding and management of the process. Hence, a mathematical model is used in this study paving the interesting intersection of biology and applied mathematics.
Methodology
- To simulate the cells’ response to the introduction of CPAs, an intricate mathematical model was created. This model incorporated the permeability of the oocyte membrane to both water and CPAs and simulated the changes occurring in the oocyte’s volume and the concentration of CPAs inside the cell during its exposure to CPA solutions.
- The results of these initial simulations were then used to develop a finite element method (FEM) model, which is a numerical technique used to find approximate solutions to boundary value problems, to capture the same responses more effectively and with spatial information.
Results & Findings
- The FEM simulations were successful in representing spatial differences during CPA permeation, like those happening near the cell membrane’s surface and those further within the cell.
- Furthermore, the FEM model was able to successfully capture changes in the shape and pressure of the cells during this CPA introduction process.
Implications
- Overall, the FEM model and simulations would allow for a more in-depth understanding of the mechanisms underlying cryopreservation outcomes. This understanding can in turn be used to optimize the cryopreservation processes used in veterinary medicine and animal husbandry, particularly in equine species.
- The study is particularly pertinent to artificial reproduction technologies, furthering their advancement and thereby improving fertility and breeding potential in animals.
Cite This Article
APA
Içli S, Soleimani M, Oldenhof H, Sieme H, Wriggers P, Wolkers WF.
(2021).
Loading equine oocytes with cryoprotective agents captured with a finite element method model.
Sci Rep, 11(1), 19812.
https://doi.org/10.1038/s41598-021-99287-9 Publication
Researcher Affiliations
- Biostabilization Laboratory - Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, NIFE, Stadtfelddamm 34, 30625, Hannover, Germany.
- Unit for Reproductive Medicine - Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany.
- Institute of Continuum Mechanics, Leibniz University Hannover, Hannover, Germany.
- Unit for Reproductive Medicine - Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany.
- Unit for Reproductive Medicine - Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany.
- Institute of Continuum Mechanics, Leibniz University Hannover, Hannover, Germany.
- Biostabilization Laboratory - Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, NIFE, Stadtfelddamm 34, 30625, Hannover, Germany. willem.frederik.wolkers@tiho-hannover.de.
- Unit for Reproductive Medicine - Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany. willem.frederik.wolkers@tiho-hannover.de.
MeSH Terms
- Animals
- Cell Membrane Permeability
- Cryopreservation / methods
- Cryoprotective Agents / pharmacology
- Horses
- Oocytes / cytology
- Vitrification
Conflict of Interest Statement
The authors declare no competing interests.
References
This article includes 27 references
- Mazur P. Freezing of living cells: mechanisms and implications.. Am J Physiol 1984 Sep;247(3 Pt 1):C125-42.
- LOVELOCK JE, BISHOP MW. Prevention of freezing damage to living cells by dimethyl sulphoxide.. Nature 1959 May 16;183(4672):1394-5.
- Fahy GM, Wowk B. Principles of cryopreservation by vitrification. Cryopreservation and Freeze-drying Protocols Methods in Molecular Biology 4. Springer; 2021. pp. 27–97.
- Wolkers WF, Oldenhof H, Tang F, Han J, Bigalk J, Sieme H. Factors Affecting the Membrane Permeability Barrier Function of Cells during Preservation Technologies.. Langmuir 2019 Jun 11;35(23):7520-7528.
- Leibo SP. Water permeability and its activation energy of fertilized and unfertilized mouse ova.. J Membr Biol 1980;53(3):179-88.
- McGrath JJ. A microscope diffusion chamber for the determination of the equilibrium and non-equilibrium osmotic response of individual cells.. J Microsc 1985 Sep;139(Pt 3):249-63.
- Gao DY, McGrath JJ, Tao J, Benson CT, Critser ES, Critser JK. Membrane transport properties of mammalian oocytes: a micropipette perfusion technique.. J Reprod Fertil 1994 Nov;102(2):385-92.
- Zhao G, Zhang Z, Zhang Y, Chen Z, Niu D, Cao Y, He X. A microfluidic perfusion approach for on-chip characterization of the transport properties of human oocytes.. Lab Chip 2017 Mar 29;17(7):1297-1305.
- KEDEM O, KATCHALSKY A. Thermodynamic analysis of the permeability of biological membranes to non-electrolytes.. Biochim Biophys Acta 1958 Feb;27(2):229-46.
- Kleinhans FW. Membrane permeability modeling: Kedem-Katchalsky vs a two-parameter formalism.. Cryobiology 1998 Dec;37(4):271-89.
- Davidson AF, Benson JD, Higgins AZ. Mathematically optimized cryoprotectant equilibration procedures for cryopreservation of human oocytes.. Theor Biol Med Model 2014 Mar 20;11:13.
- Anderson DM, Benson JD, Kearsley AJ. Foundations of modeling in cryobiology-III: Inward solidification of a ternary solution towards a permeable spherical cell in the dilute limit.. Cryobiology 2020 Feb 1;92:34-46.
- Anderson DM, Benson JD, Kearsley AJ. Foundations of modeling in cryobiology-II: Heat and mass transport in bulk and at cell membrane and ice-liquid interfaces.. Cryobiology 2019 Dec;91:3-17.
- Kojic M, Milosevic M, Simic V, Koay EJ, Kojic N, Ziemys A, Ferrari M. Multiscale smeared finite element model for mass transport in biological tissue: From blood vessels to cells and cellular organelles.. Comput Biol Med 2018 Aug 1;99:7-23.
- Hansen KB, Arzani A, Shadden SC. Finite element modeling of near-wall mass transport in cardiovascular flows.. Int J Numer Method Biomed Eng 2019 Jan;35(1):e3148.
- Hajiyavand AM, Saadat M, Abena A, Sadak F, Sun X. Effect of Injection Speed on Oocyte Deformation in ICSI.. Micromachines (Basel) 2019 Mar 29;10(4).
- Hajiyavand AM, Saadat M, Stamboulis A. A mechanical model for an artificial oocyte. Int. J. Model. Optim. 2017;7:315–321.
- Lotz J, Içli S, Liu D, Caliskan S, Sieme H, Wolkers WF, Oldenhof H. Transport processes in equine oocytes and ovarian tissue during loading with cryoprotective solutions.. Biochim Biophys Acta Gen Subj 2021 Feb;1865(2):129797.
- Lahmann JM, Sanchez CC, Benson JD, Acker JP, Higgins AZ. Implications of variability in cell membrane permeability for design of methods to remove glycerol from frozen-thawed erythrocytes.. Cryobiology 2020 Feb 1;92:168-179.
- Zhao G, Fu J. Microfluidics for cryopreservation.. Biotechnol Adv 2017 Mar-Apr;35(2):323-336.
- Zhurova M, Lusianti RE, Higgins AZ, Acker JP. Osmotic tolerance limits of red blood cells from umbilical cord blood.. Cryobiology 2014 Aug;69(1):48-54.
- De Coster T, Velez DA, Van Soom A, Woelders H, Smits K. Cryopreservation of equine oocytes: looking into the crystal ball.. Reprod Fertil Dev 2020 Mar;32(5):453-467.
- Ducheyne KD, Rizzo M, Daels PF, Stout TAE, de Ruijter-Villani M. Vitrifying immature equine oocytes impairs their ability to correctly align the chromosomes on the MII spindle.. Reprod Fertil Dev 2019 Jul;31(8):1330-1338.
- Mullen SF, Rosenbaum M, Critser JK. The effect of osmotic stress on the cell volume, metaphase II spindle and developmental potential of in vitro matured porcine oocytes.. Cryobiology 2007 Jun;54(3):281-9.
- Dill E. The finite element method for mechanics of solids with ANSYS applications. CRC Press; 2011.
- Boccaccio A, Frassanito MC, Lamberti L, Brunelli R, Maulucci G, Monaci M, Papi M, Pappalettere C, Parasassi T, Sylla L, Ursini F, De Spirito M. Nanoscale characterization of the biomechanical hardening of bovine zona pellucida.. J R Soc Interface 2012 Nov 7;9(76):2871-82.
- Zieger MA, Tredget EE, Sykes BD, McGann LE. Injury and protection in split-thickness skin after very rapid cooling and warming.. Cryobiology 1997 Aug;35(1):53-69.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists