Separating the contributions of zona pellucida and cytoplasm in the viscoelastic response of human oocytes.

The successful characterization of the mechanical properties of human oocytes and young embryos is of crucial relevance to reduce the risk of pregnancy arrest in in-vitro fertilization processes. Unfortunately, current study has been hindered by the lack of accuracy in describing the mechanical contributions of each structure (zona pellucida, cytoplasm) due to its high heterogeneity. In this work, we present a novel approach to model the oocyte response taking into account the effect of both zona and cytoplasm, as well as different loading conditions. The model is then applied to develop an experimental protocol capable of accurately separating the viscoelastic contribution of zona and cytoplasm by simply varying the loading condition. This new protocol has the potential to open the door to improving our understanding the mechanical properties of oocytes at different stages, and provide a quantitative predictive ability to the evaluation of oocyte quality. STATEMENT OF SIGNIFICANCE: Assisted reproductive technologies, such as in vitro fertilization, often rely on identifying high quality oocytes or female egg cells. The viscoelastic properties of these cells, such as stiffness and stress relaxation time, have been identified as potential objective indicators of cell quality. However, their characterization has proven difficult due to the structural heterogeneity of the cell and inconsistent loading conditions. This paper presents a new model that, although simple, addresses the above difficulties to provide accurate estimations of the cell's mechanical properties. Learning from this model, we then propose a novel non-invasive testing protocol to allow oocyte characterization with increased accuracy. We believe this effort would improve consistency in measurements and enhance our knowledge on the mechanics of oocytes.

[1]  Markus Böl,et al.  Mechanical characterisation of oocytes - The influence of sample geometry on parameter identification. , 2018, Journal of the mechanical behavior of biomedical materials.

[2]  Chang Kyoung Choi,et al.  Simultaneous measurements of cytoplasmic viscosity and intracellular vesicle sizes for live human brain cancer cells. , 2011, Biotechnology and bioengineering.

[3]  S. Omata,et al.  Micro-mechanical sensing platform for the characterization of the elastic properties of the ovum via uniaxial measurement. , 2004, Journal of biomechanics.

[4]  Dag Normann,et al.  Mathematics of Computation at CiE 2005 , 2006 .

[5]  Massimiliano Papi,et al.  Viscous forces are predominant in the zona pellucida mechanical resistance , 2013 .

[6]  Byungkyu Kim,et al.  Cell Stiffness Is a Biomarker of the Metastatic Potential of Ovarian Cancer Cells , 2012, PloS one.

[7]  Geraldine Seydoux,et al.  Regulation of the Oocyte-to-Zygote Transition , 2007, Science.

[8]  P Zioupos,et al.  Mechanical properties and the hierarchical structure of bone. , 1998, Medical engineering & physics.

[9]  Franck J. Vernerey,et al.  A mathematical model of the coupled mechanisms of cell adhesion, contraction and spreading , 2013, Journal of Mathematical Biology.

[10]  Robert P. Edwards Reproductive BioMedicine Online is moving to 12 issues per annum , 2004 .

[11]  Franck J. Vernerey A theoretical treatment on the mechanics of interfaces in deformable porous media , 2011 .

[12]  Horn-Sen Tzou,et al.  SENSORS AND ACTUATORS , 2001 .

[13]  Kwang W. Jeon,et al.  International review of cell and molecular biology , 2008 .

[14]  K. Seffen,et al.  International Journal of Solids and Structures , 2015 .

[15]  Luciano Lamberti,et al.  Effect of AFM probe geometry on visco-hyperelastic characterization of soft materials , 2015, Nanotechnology.

[16]  M. Papi,et al.  Mechanical properties of zona pellucida hardening , 2010, European Biophysics Journal.

[17]  Basak Balaban,et al.  Effect of oocyte morphology on embryo development and implantation. , 2006, Reproductive biomedicine online.

[18]  Williams,et al.  Compressive deformation of a single microcapsule. , 1996, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[19]  A. Peirce Computer Methods in Applied Mechanics and Engineering , 2010 .

[20]  P. Wassarman,et al.  Constructing the mammalian egg zona pellucida: some new pieces of an old puzzle. , 1996, Journal of cell science.

[21]  R. Williams,et al.  Journal of American Chemical Society , 1979 .

[22]  M. Navidbakhsh,et al.  Alteration in the Mechanical Properties of Human Ovum Zona Pellucida Following Fertilization: Experimental and Analytical Studies , 2011 .

[23]  Yu Sun,et al.  In-situ mechanical characterization of mouse oocytes using a cell holding device , 2010, 2010 IEEE 23rd International Conference on Micro Electro Mechanical Systems (MEMS).

[24]  W SEELENTAG [RADIATION AND ENVIRONMENT]. , 1964, Rontgenpraxis; Zeitschrift fur radiologische Technik.

[25]  D F Katz,et al.  Biophysical properties of the zona pellucida measured by capillary suction: is zona hardening a mechanical phenomenon? , 1988, The Journal of experimental zoology.

[26]  Sadao Omata,et al.  Mouse zona pellucida dynamically changes its elasticity during oocyte maturation, fertilization and early embryo development , 2006, Human Cell.

[27]  Z. Rosenwaks,et al.  Oolemma characteristics in relation to survival and fertilization patterns of oocytes treated by intracytoplasmic sperm injection. , 1996, Human reproduction.

[28]  P A Valberg,et al.  Magnetic particle motions within living cells. Measurement of cytoplasmic viscosity and motile activity. , 1987, Biophysical journal.

[29]  H. Keller,et al.  Analysis of Numerical Methods , 1967 .

[30]  Yu Sun,et al.  Elastic and Viscoelastic Characterization of Mouse Oocytes Using Micropipette Indentation , 2012, Annals of Biomedical Engineering.

[31]  Martin Lenz,et al.  Engineering Elasticity and Relaxation Time in Metal-Coordinate Cross-Linked Hydrogels , 2016 .

[32]  O. Thoumine,et al.  Time scale dependent viscoelastic and contractile regimes in fibroblasts probed by microplate manipulation. , 1997, Journal of cell science.

[33]  K. Jacobson,et al.  Local measurements of viscoelastic parameters of adherent cell surfaces by magnetic bead microrheometry. , 1998, Biophysical journal.

[34]  A S Verkman,et al.  Low viscosity in the aqueous domain of cell cytoplasm measured by picosecond polarization microfluorimetry , 1991, The Journal of cell biology.

[35]  C. Lim,et al.  Biomechanics approaches to studying human diseases. , 2007, Trends in biotechnology.

[36]  B D Slenning,et al.  Agreement among evaluators of bovine embryos produced in vivo or in vitro. , 1995, Theriogenology.

[37]  K Luby-Phelps,et al.  Viscoelastic response of fibroblasts to tension transmitted through adherens junctions. , 1997, Biophysical journal.

[38]  Norbert Gleicher Journal of Assisted Reproduction and Genetics 2002 Reviewers , 2004, Journal of Assisted Reproduction and Genetics.

[39]  P A Valberg,et al.  Cytoplasmic motions, rheology, and structure probed by a novel magnetic particle method , 1985, The Journal of cell biology.

[40]  J. Toca-Herrera,et al.  Stress relaxation microscopy: imaging local stress in cells. , 2010, Journal of biomechanics.

[41]  S. Kahraman,et al.  Relationship between granular cytoplasm of oocytes and pregnancy outcome following intracytoplasmic sperm injection. , 2000, Human reproduction.

[42]  Iku Nemoto,et al.  A Model of Magnetization and Relaxation of Ferrimagnetic Particles in the Lung , 1982, IEEE Transactions on Biomedical Engineering.

[43]  R M Nerem,et al.  Viscoelastic properties of cultured porcine aortic endothelial cells exposed to shear stress. , 1996, Journal of biomechanics.

[44]  R. Waugh,et al.  Shear Rate-Dependence of Leukocyte Cytoplasmic Viscosity , 1994 .

[45]  Denis Wirtz,et al.  Probing single-cell micromechanics in vivo: the microrheology of C. elegans developing embryos. , 2006, Biophysical journal.

[46]  M. Radmacher,et al.  Substrate dependent differences in morphology and elasticity of living osteoblasts investigated by atomic force microscopy. , 2000, Colloids and surfaces. B, Biointerfaces.

[47]  Nature Protocols , 2006 .

[48]  R. Edwards,et al.  Reproductive BioMedicine Online accepted for Impact Factor. , 2005, Reproductive biomedicine online.

[49]  P Xia,et al.  Intracytoplasmic sperm injection: correlation of oocyte grade based on polar body, perivitelline space and cytoplasmic inclusions with fertilization rate and embryo quality. , 1997, Human reproduction.

[50]  Transport in Porous Media Contents of Volume 46 , 2002 .

[51]  S Chien,et al.  Leukocyte relaxation properties. , 1988, Biophysical journal.

[52]  G. Semenza,et al.  Measuring elasticity of biological materials by atomic force microscopy , 1998, FEBS letters.

[53]  J. Herskowitz,et al.  Proceedings of the National Academy of Sciences, USA , 1996, Current Biology.

[54]  S. Bryant,et al.  Tuning tissue growth with scaffold degradation in enzyme-sensitive hydrogels: a mathematical model. , 2016, Soft matter.

[55]  E. Sackmann,et al.  Measurement of local viscoelasticity and forces in living cells by magnetic tweezers. , 1999, Biophysical journal.

[56]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.

[57]  S. Saravananb,et al.  Colloids and Surfaces B: Biointerfaces , 2019 .

[58]  Lar,et al.  Reproduction , 1975, Comprehensive Virology.

[59]  Haimin Yao,et al.  Journal of the Mechanics and Physics of Solids , 2014 .

[60]  Tsuyoshi Murata,et al.  {m , 1934, ACML.

[61]  M Sommergruber,et al.  Developmental competence of oocytes showing increased cytoplasmic viscosity. , 2003, Human reproduction.

[62]  E. Evans,et al.  Apparent viscosity and cortical tension of blood granulocytes determined by micropipet aspiration. , 1989, Biophysical journal.

[63]  E. Triphosphat,et al.  FEBS Letters , 1987, FEBS Letters.

[64]  G B Schuessler,et al.  Influence of physicochemical factors on rheology of human neutrophils. , 1982, Biophysical journal.

[65]  Jaromír Slavík,et al.  COMPUTATIONAL MECHANICS I , 2004 .

[66]  Michael S Sacks,et al.  Design and analysis of tissue engineering scaffolds that mimic soft tissue mechanical anisotropy. , 2006, Biomaterials.

[67]  E. G. Jones Cerebral Cortex , 1987, Cerebral Cortex.