European Biophysics Journal

, Volume 48, Issue 6, pp 585–592 | Cite as

Study of the mechanical properties of fresh and cryopreserved individual human oocytes

  • Elena Giolo
  • Monica Martinelli
  • Stefania Luppi
  • Federico Romano
  • Giuseppe RicciEmail author
  • Marco Lazzarino
  • Laura AndolfiEmail author
Biophysics Letter


In assisted reproduction technologies, the cryopreservation of oocytes is a common procedure used to circumvent female infertility. However, some morphological and functional alterations of oocytes have been observed depending on the protocol applied. In this work, the mechanical response of individual human oocytes before and after a freeze-thawing procedure was characterised. Oocytes, immediately after retrieval, were morphologically evaluated by bright-field optical microscopy and their elasticity measured by indentation measurements using atomic force microscopy. Oocytes were then frozen according to the open-vitrification protocol and stored in liquid nitrogen. Afterwards, the same oocytes were thawed and the indentation measurements repeated. Using this approach, we can follow the elasticity of a set of single oocytes from retrieval up to the freeze-thawing procedure. The analysis of the resulting data shows that the retrieved healthy oocytes, which preserve their healthy morphological features after cryopreservation, maintain unchanged also in stiffness values. In contrast, oocytes having dysmorphic characteristics, before and/or after freeze-thawing, show significant variations in their mechanical response. In addition, the dysmorphic oocytes are generally observed to be softer than the healthy oocytes. Our results indicate that stiffness of healthy oocytes is not considerably affected by the open-vitrification-thawing procedure, and that distinct elasticity ranges can be identified for healthy and dysmorphic oocytes. These findings indicate that the mechanical characterization of oocytes represents an opportunity to detect cellular defects, and assess the quality and bio-viability of processes such as cryopreservation.


Oocytes Vitrification Atomic Force Microscopy Biomechanics In vitro fertilization 



This work was supported by Health Ministry (RF-2011–02351812) Ricerca Finalizzata “Clinical Applications of Ultrastructural Cell Analysis in the Field of Reproductive Technologies” and by Regione Friuli Venezia Giulia, within the framework of “Regional Law 17/2004: Contributions for clinical, translational, basic, epidemiological and organizational research”, with the project “BioMec—Application of biomechanical technologies to integrate traditional methods in the hospital context”. We thank Simone dal Zilio for PDMS supports and Ines Delfino for helping with the Matlab routine.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Andolfi L, Bourkoula E, Migliorini E, Palma A, Pucer A, Skrap M, Scoles G, Beltrami AP, Cesselli D, Lazzarino M (2014) Investigation of Adhesion and Mechanical Properties of Human Glioma Cells by Single Cell Force Spectroscopy and Atomic Force Microscopy. PLoS ONE 9:e112582CrossRefGoogle Scholar
  2. Andolfi L, Masiero E, Giolo E, Martinelli M, Luppi S, Dal Zilio S, Delfino I, Bortul R, Zweyer M, Ricci G, Lazzarino M (2017) Investigating the mechanical properties of zona pellucida of whole human oocytes by atomic force spectroscopy. Integr Biol 8:886–893CrossRefGoogle Scholar
  3. Argyle CE, Harper JC, Davies MC (2016) Oocyte cryopreservation: where are we now? Hum Reprod Update 22:440–449CrossRefGoogle Scholar
  4. Bianchi V, Macchiarelli G, Borini A, Lappi M, Cecconi S, Miglietta S, Familiari G, Nottola SA (2014) Fine morphological assessment of quality of human mature oocytes after slow freezing or vitrification with a closed device: a comparative analysis. Reprod Biol Endocrinol 12:110–2331CrossRefGoogle Scholar
  5. Choi JK, Yue T, Huang H, Zhao G, Zhang M, He X (2015) The crucial role of zona pellucida in cryopreservation of oocytes by vitrification. Cryobiology 71:350–355CrossRefGoogle Scholar
  6. Cross SE, Jin Y-S, Rao J, Gimzewski JK (2007) Nanomechanical analysis of cells from cancer patients. Nat Nanotech 2:780–783CrossRefGoogle Scholar
  7. Ghetler Y, Skutelsky E, Ben Nun I, Ben Dor L, Amihai D, Shalgi R ((2006) ) Human oocyte cryopreservation and the fate of cortical granules. Journal of Ecology 86:210–216–216CrossRefGoogle Scholar
  8. Glujovsky D, Riestra B, Sueldo C, Fiszbajn G, Repping S, Nodar F, Papier S, Ciapponi A. (2014) Vitrification versus slow freezing for women undergoing oocyte cryopreservation. Cochrane Database Syst Rev 9: CD010047.Google Scholar
  9. Gu R, Feng Y, Guo S, Zhao S, Lu X, Fu J, Sun X, Sun Y Cryobiology 75:144–150CrossRefGoogle Scholar
  10. Islam M, Brink H, Blanche S, DiPrete C, Bongiorno T, Stone N, Liu A, Philip A, Wang G, Lam W, Alexeev A, Waller EK, Sulchek T (2017) Microfluidic, Sorting of Cells by Viability Based on Differences in Cell Stiffness. Sci Rep 7:1997–859CrossRefGoogle Scholar
  11. Khalilian M, Navidbakhsh M, Valojerdi MR, Chizari M, Yazdi PE (2010) Estimating Young’s modulus of zona pellucida by micropipette aspiration in combination with theoretical models of ovum. J R Soc Interface 7:687–694CrossRefGoogle Scholar
  12. Kort J, Behr B (2015) When maladaptive gene flow does not increase selection. Evolution 69:2289–741CrossRefGoogle Scholar
  13. Ledda S, Bogliolo L, Succu S, Ariu F, Bebbere D, Leoni GG, Naitana S (2003) Contemporary evolution meets conservation biology. Trends in Ecology & Evolution 18:94CrossRefGoogle Scholar
  14. Murayama Y, Mizuno J, Kamakura H, Fueta Y, Nakamura H, Akaishi K, Anzai K, Watanabe A, Inui H, Omata S (2016) High fitness costs of climate change-induced camouflage mismatchHum Cell 19:119–125CrossRefGoogle Scholar
  15. Nguyen AV, Nyberg KD, Scott MB, Welsh AM, Nguyen AH, Wu N, Hohlbauch SV, Geisse NA, Gibb EA, Robertson AG, Donahued TR, Rowat AC (2016) Stiffness of pancreatic cancer cells is associated with increased invasive potential. Integr Biol 12:1232–1245CrossRefGoogle Scholar
  16. Otto O, Rosendahl P, Mietke A, Golfier S, Herold C, Klaue D, Girardo S, Pagliara S, Ekpenyong A, Jacobi A, Wobus M, Töpfner N, Keyser UF, Mansfeld J, Fischer-Friedrich E, Guck J (2015) Real-time deformability cytometry: on-the-fly cell mechanical phenotyping. Nat Methods 12:199–202CrossRefGoogle Scholar
  17. Papi M, Brunelli R, Sylla L, Parasassi T, Monaci M, Maulucci G, Missori M, Arcovito G, Ursini F, De Spirito M (2010) Mechanical properties of zona pellucida hardening. Eur Biophys J 39:987–992CrossRefGoogle Scholar
  18. Plodinec M, Loparic M, Monnier CA, Obermann EC, Zanetti-Dallenbach R, Oertle P, Hyotyla JT, Aebi U, Bentires-Alj M, Lim RYH, Schoenenberger CA (2012) The nanomechanical signature of breast cancer. Nat Nanotech 7:757–765CrossRefGoogle Scholar
  19. Ricci G, Granzotto M, Luppi S, Giolo E, Martinelli M, Zito G, Borelli M (2015) Effect of seminal leukocytes on in vitro fertilization and intracytoplasmic sperm injection outcomes. Fertil Steril 104:87–93CrossRefGoogle Scholar
  20. Rienzi L, Vajta G, Ubaldi F (2011) Predictive value of oocyte morphology in human IVF: a systematic review of the literature. Hum Reprod Update 17:34–45CrossRefGoogle Scholar
  21. Rusciano G, De Canditiis C, Zito G, Rubessa M, Roca MS, Carotenuto R, Sasso A, Gasparrini B (2017) Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes. PLoS ONE 12:e0177677CrossRefGoogle Scholar
  22. Saragusty J, Arav A (2011) Current progress in oocyte and embryo cryopreservation by slow freezing and vitrification. Reproduction 141:1–19CrossRefGoogle Scholar
  23. Stoop D (2010) Social oocyte freezing. FV & V in ObGyn 2:31–34Google Scholar
  24. Sun Y, Wan K-T, Roberts K-P, Bischof JP, Nelson BJ (2003) Mechanical Property Characterization of Mouse Zona Pellucida. IEEE Trans on Nanobioscience. 2:279–286CrossRefGoogle Scholar
  25. Te Rieta J, Katanc AJ, Rankl C, Stahl SW, Van Buul AM, Phang IY, Gomez-Casado A, Schöng P, Gerritsen JW, Cambi A, Rowan AE, Vancso GJ, Jonkheijm P, Huskens J, Oosterkamp TH, Gaub H, Hinterdorfer P, Figdor CG, Speller S (2011) Interlaboratory round robin on cantilever calibration for AFM force spectroscopy. Ultramicroscopy 111:1659–1669CrossRefGoogle Scholar
  26. Yanez LZ, Camarillo DB (2017) Microfluidic analysis of oocyte and embryo biomechanical properties to improve outcomes in assisted reproductive technologies Molec. Hum Reprod 23:235–247CrossRefGoogle Scholar
  27. Yanez LZ, Han J, Behr BB, Reijo Pera RA, Camarillo DB (2016) Human oocyte developmental potential is predicted by mechanical properties within hours after fertilization. Nat Commun 7:10809CrossRefGoogle Scholar
  28. Zemła J, Danilkiewicz J, Orzechowska B, Pabijan J, Seweryn S, Lekka M (2018) Atomic force microscopy as a tool for assessing the cellular elasticity and adhesiveness to identify cancer cells and tissues. Semin Cell Dev Biol 73:115–124CrossRefGoogle Scholar
  29. Zenzes MT, Bielecki R, Casper RF, Leibo SP (2001) Effects of chilling to 0 degrees C on the morphology of meiotic spindles in human metaphase II oocytes. Fertil Steril 75:769–777CrossRefGoogle Scholar

Copyright information

© European Biophysical Societies' Association 2019

Authors and Affiliations

  1. 1.Institute for Maternal and Child HealthIRCCS Burlo GarofoloTriesteItaly
  2. 2.Department of Medicine, Surgery and Health SciencesUniversity of TriesteTriesteItaly
  3. 3.Istituto Officina dei Materiali, Consiglio Nazionale delle RicercheTriesteItaly

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