High pressure frozen oocytes have improved ultrastructure but reduced cleavage rates compared to conventionally fixed or vitrified oocytes
K. L. Reader
A * , B. G. Pilbrow B , S. Zellhuber-McMillan A , A. J. Mitchell C , J. L. Juengel D and D. Morbeck E
+ Author Affiliations
- Author Affiliations
A Department of Pathology, University of Otago, Dunedin 9016, New Zealand.
B Department of Anatomy, University of Otago, Dunedin 9016, New Zealand.
C Otago Micro and Nanoscale Imaging Electron Microscopy Unit, University of Otago, Dunedin 9016, New Zealand.
D AgResearch, Invermay Agricultural Centre, Puddle Alley, Mosgiel, New Zealand.
E Fertility Associates, Auckland, New Zealand.
Reproduction, Fertility and Development 34(18) 1135-1144 https://doi.org/10.1071/RD22118
Published online: 2 November 2022
© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing
Abstract
Context: Live birth rates are lower for cryopreserved oocytes than for fresh IVF cycles, indicating a need for improved methodologies.
Aims: The aim of this study was to determine if high pressure freezing (HPF) could improve both ultrastructural preservation and cryopreserved oocyte quality when compared to conventional fixation and vitrification methods.
Methods: Sheep oocytes and embryos were prepared by HPF or vitrification, with or without cryoprotectants. Frozen oocytes were prepared for transmission electron microscopy or warmed, in vitro fertilised and the recovery and cleavage rates recorded.
Key results: Blastocyst rates were similar between fresh, HPF and vitrified embryos. HPF oocytes had improved ultrastructure compared to conventional fixation or vitrification, but had poorer survival and cleavage rates compared to vitrified oocytes. Freeze-substitution of cryopreserved oocytes and transmission electron microscopy demonstrated disruption of the oocyte ultrastructure in the presence of cryoprotectants.
Conclusions: Superior preservation of ultrastructure was observed in HPF oocytes compared to vitrification or conventional fixation methods. In the presence of CP, both embryos and oocytes could survive HPF and warming but oocytes had reduced development.
Implications: The HPF method has potential to be developed and lead to improved oocyte and embryo cryopreservation and outcomes for assisted reproduction.
Keywords: cryopreservation, embryo, high pressure freezing, in vitro maturation, oocyte, transmission electron microscopy, ultrastructure, vitrification.
References
Arav, A (2014). Cryopreservation of oocytes and embryos. Theriogenology 81, 96–102.| Cryopreservation of oocytes and embryos.Crossref | GoogleScholarGoogle Scholar |
Asgari, V, Hosseini, SM, Ostadhosseini, S, Hajian, M, and Nasr-Esfahani, MH (2011). Time dependent effect of post warming interval on microtubule organization, meiotic status, and parthenogenetic activation of vitrified in vitro matured sheep oocytes. Theriogenology 75, 904–910.
| Time dependent effect of post warming interval on microtubule organization, meiotic status, and parthenogenetic activation of vitrified in vitro matured sheep oocytes.Crossref | GoogleScholarGoogle Scholar |
Azari, M, Kafi, M, Ebrahimi, B, Fatehi, R, and Jamalzadeh, M (2017). Oocyte maturation, embryo development and gene expression following two different methods of bovine cumulus-oocyte complexes vitrification. Veterinary Research Communications 41, 49–56.
| Oocyte maturation, embryo development and gene expression following two different methods of bovine cumulus-oocyte complexes vitrification.Crossref | GoogleScholarGoogle Scholar |
Bhattacharya, S, Hamilton, MPR, Shaaban, M, Khalaf, Y, Seddler, M, Ghobara, T, Braude, P, Kennedy, R, Rutherford, A, Hartshorne, G, and Templeton, A (2001). Conventional in-vitro fertilisation versus intracytoplasmic sperm injection for the treatment of non-male-factor infertility: a randomised controlled trial. The Lancet 357, 2075–2079.
| Conventional in-vitro fertilisation versus intracytoplasmic sperm injection for the treatment of non-male-factor infertility: a randomised controlled trial.Crossref | GoogleScholarGoogle Scholar |
Bogliolo, L, Ledda, S, Innocenzi, P, Ariu, F, Bebbere, D, Rosati, I, Leoni, GG, and Piccinini, M (2012). Raman microspectroscopy as a non-invasive tool to assess the vitrification-induced changes of ovine oocyte zona pellucida. Cryobiology 64, 267–272.
| Raman microspectroscopy as a non-invasive tool to assess the vitrification-induced changes of ovine oocyte zona pellucida.Crossref | GoogleScholarGoogle Scholar |
Boulet, SL, Mehta, A, Kissin, DM, Warner, L, Kawwass, JF, and Jamieson, DJ (2015). Trends in use of and reproductive outcomes associated with intracytoplasmic sperm injection. JAMA 313, 255–263.
| Trends in use of and reproductive outcomes associated with intracytoplasmic sperm injection.Crossref | GoogleScholarGoogle Scholar |
Chian, R-C, Uzelac, PS, and Nargund, G (2013). In vitro maturation of human immature oocytes for fertility preservation. Fertility and Sterility 99, 1173–1181.
| In vitro maturation of human immature oocytes for fertility preservation.Crossref | GoogleScholarGoogle Scholar |
Davies, MJ, Moore, VM, Willson, KJ, Van Essen, P, Priest, K, Scott, H, Haan, EA, and Chan, A (2012). Reproductive technologies and the risk of birth defects. New England Journal of Medicine 366, 1803–1813.
| Reproductive technologies and the risk of birth defects.Crossref | GoogleScholarGoogle Scholar |
dos Santos-Neto, PC, Vilariño, M, Cuadro, F, Barrera, N, Crispo, M, and Menchaca, A (2020). Cumulus cells during in vitro fertilization and oocyte vitrification in sheep: remove, maintain or add? Cryobiology 92, 161–167.
| Cumulus cells during in vitro fertilization and oocyte vitrification in sheep: remove, maintain or add?Crossref | GoogleScholarGoogle Scholar |
Fasano, G, Demeestere, I, and Englert, Y (2012). In-vitro maturation of human oocytes: before or after vitrification? Journal of Assisted Reproduction and Genetics 29, 507–512.
| In-vitro maturation of human oocytes: before or after vitrification?Crossref | GoogleScholarGoogle Scholar |
Gjerris, AC, Loft, A, Pinborg, A, Christiansen, M, and Tabor, A (2008). Prenatal testing among women pregnant after assisted reproductive techniques in Denmark 1995–2000: a national cohort study. Human Reproduction 23, 1545–1552.
| Prenatal testing among women pregnant after assisted reproductive techniques in Denmark 1995–2000: a national cohort study.Crossref | GoogleScholarGoogle Scholar |
Goldman, KN, Noyes, NL, Knopman, JM, McCaffrey, C, and Grifo, JA (2013). Oocyte efficiency: does live birth rate differ when analyzing cryopreserved and fresh oocytes on a per-oocyte basis? Fertility and Sterility 100, 712–717.
| Oocyte efficiency: does live birth rate differ when analyzing cryopreserved and fresh oocytes on a per-oocyte basis?Crossref | GoogleScholarGoogle Scholar |
Gook, DA, and Edgar, DH (2007). Human oocyte cryopreservation. Human Reproduction Update 13, 591–605.
| Human oocyte cryopreservation.Crossref | GoogleScholarGoogle Scholar |
Gualtieri, R, Mollo, V, Barbato, V, Fiorentino, I, Iaccarino, M, and Talevi, R (2011). Ultrastructure and intracellular calcium response during activation in vitrified and slow-frozen human oocytes. Human Reproduction 26, 2452–2460.
| Ultrastructure and intracellular calcium response during activation in vitrified and slow-frozen human oocytes.Crossref | GoogleScholarGoogle Scholar |
Hosseini, SM, Asgari, V, Ostadhosseini, S, Hajian, M, Piryaei, A, Najarasl, M, and Nasr-Esfahani, MH (2012). Potential applications of sheep oocytes as affected by vitrification and in vitro aging. Theriogenology 77, 1741–1753.
| Potential applications of sheep oocytes as affected by vitrification and in vitro aging.Crossref | GoogleScholarGoogle Scholar |
Hosseini, SM, Asgari, V, Ostadhosseini, S, Hajian, M, Ghanaei, HR, and Nasr-Esfahani, MH (2015). Developmental competence of ovine oocytes after vitrification: differential effects of vitrification steps, embryo production methods, and parental origin of pronuclei. Theriogenology 83, 366–376.
| Developmental competence of ovine oocytes after vitrification: differential effects of vitrification steps, embryo production methods, and parental origin of pronuclei.Crossref | GoogleScholarGoogle Scholar |
Huebinger, J, Han, H-M, and Grabenbauer, M (2016). Reversible cryopreservation of living cells using an electron microscopy cryo-fixation method. PLoS ONE 11, e0164270.
| Reversible cryopreservation of living cells using an electron microscopy cryo-fixation method.Crossref | GoogleScholarGoogle Scholar |
Hwang, J-L, Lin, Y-H, and Tsai, Y-L (2000). In vitro maturation and fertilization of immature oocytes: a comparative study of fertilization techniques. Journal of Assisted Reproduction and Genetics 17, 39–43.
| In vitro maturation and fertilization of immature oocytes: a comparative study of fertilization techniques.Crossref | GoogleScholarGoogle Scholar |
Kang, M-H, You, S-Y, Hong, K, and Kim, J-H (2020). DMSO impairs the transcriptional program for maternal-to-embryonic transition by altering histone acetylation. Biomaterials 230, 119604.
| DMSO impairs the transcriptional program for maternal-to-embryonic transition by altering histone acetylation.Crossref | GoogleScholarGoogle Scholar |
Kopeika, J, Thornhill, A, and Khalaf, Y (2015). The effect of cryopreservation on the genome of gametes and embryos: principles of cryobiology and critical appraisal of the evidence. Human Reproduction Update 21, 209–227.
| The effect of cryopreservation on the genome of gametes and embryos: principles of cryobiology and critical appraisal of the evidence.Crossref | GoogleScholarGoogle Scholar |
Kushnir, VA, Darmon, SK, Barad, DH, and Gleicher, N (2018). New national outcome data on fresh versus cryopreserved donor oocytes. Journal of Ovarian Research 11, 2.
| New national outcome data on fresh versus cryopreserved donor oocytes.Crossref | GoogleScholarGoogle Scholar |
Lane, M, Robker, RL, and Robertson, SA (2014). Parenting from before conception. Science 345, 756–760.
| Parenting from before conception.Crossref | GoogleScholarGoogle Scholar |
Leibo, SP, and Pool, TB (2011). The principal variables of cryopreservation: solutions, temperatures, and rate changes. Fertility and Sterility 96, 269–276.
| The principal variables of cryopreservation: solutions, temperatures, and rate changes.Crossref | GoogleScholarGoogle Scholar |
Manning L, Richmond J (2015) High-pressure freeze and freeze substitution electron microscopy in C. elegans. In ‘C. elegans: methods and applications’. (Eds D Biron, G Haspel) pp. 121–140. (Humana Press: Totowa, NJ)
McDonald, KL, and Auer, M (2006). High-pressure freezing, cellular tomography, and structural cell biology. BioTechniques 41, 137–143.
| High-pressure freezing, cellular tomography, and structural cell biology.Crossref | GoogleScholarGoogle Scholar |
Nagy, ZP, Nel-Themaat, L, Chang, C-C, Shapiro, DB, and Berna, DP (2014). Cryopreservation of eggs. Methods in Molecular Biology 1154, 439–454.
| Cryopreservation of eggs.Crossref | GoogleScholarGoogle Scholar |
Nixon, SJ, Webb, RI, Floetenmeyer, M, Schieber, N, Lo, HP, and Parton, RG (2009). A single method for cryofixation and correlative light, electron microscopy and tomography of zebrafish embryos. Traffic 10, 131–136.
| A single method for cryofixation and correlative light, electron microscopy and tomography of zebrafish embryos.Crossref | GoogleScholarGoogle Scholar |
Nottola, SA, Coticchio, G, Sciajno, R, Gambardella, A, Maione, M, Scaravelli, G, Bianchi, S, Macchiarelli, G, and Borini, A (2009). Ultrastructural markers of quality in human mature oocytes vitrified using cryoleaf and cryoloop. Reproductive BioMedicine Online 19, 17–27.
| Ultrastructural markers of quality in human mature oocytes vitrified using cryoleaf and cryoloop.Crossref | GoogleScholarGoogle Scholar |
Potdar, N, Gelbaya, TA, and Nardo, LG (2014). Oocyte vitrification in the 21st century and post-warming fertility outcomes: a systematic review and meta-analysis. Reproductive BioMedicine Online 29, 159–176.
| Oocyte vitrification in the 21st century and post-warming fertility outcomes: a systematic review and meta-analysis.Crossref | GoogleScholarGoogle Scholar |
Practice Committees of the American Society for Reproductive Medicine and the Society for Assisted Reproductive Technology (2020). Intracytoplasmic sperm injection (ICSI) for non–male factor indications: a committee opinion. Fertility and Sterility 114, 239–245.
| Intracytoplasmic sperm injection (ICSI) for non–male factor indications: a committee opinion.Crossref | GoogleScholarGoogle Scholar |
Reader, KL, Cox, NR, Stanton, J-AL, and Juengel, JL (2015). Mitochondria and vesicles differ between adult and prepubertal sheep oocytes during IVM. Reproduction, Fertility and Development 27, 513–522.
| Mitochondria and vesicles differ between adult and prepubertal sheep oocytes during IVM.Crossref | GoogleScholarGoogle Scholar |
Sandin, S, Nygren, K-G, Iliadou, A, Hultman, CM, and Reichenberg, A (2013). Autism and mental retardation among offspring born after in vitro fertilization. JAMA 310, 75–84.
| Autism and mental retardation among offspring born after in vitro fertilization.Crossref | GoogleScholarGoogle Scholar |
Seki, S, and Mazur, P (2012). Ultra-rapid warming yields high survival of mouse oocytes cooled to −196°C in dilutions of a standard vitrification solution. PLoS ONE 7, e36058.
| Ultra-rapid warming yields high survival of mouse oocytes cooled to −196°C in dilutions of a standard vitrification solution.Crossref | GoogleScholarGoogle Scholar |
The European IVF-Monitoring Consortium (EIM) for the European Society of Human Reproduction and Embryology (ESHRE) Wyns, C, De Geyter, C, Calhaz-Jorge, C, Kupka, MS, Motrenko, T, Smeenk, J, Bergh, C, Tandler-Schneider, A, Rugescu, IA, Vidakovic, S, and Goossens, V (2021). ART in Europe, 2017: results generated from European registries by ESHRE. Human Reproduction Open 2021, hoab026.
| ART in Europe, 2017: results generated from European registries by ESHRE.Crossref | GoogleScholarGoogle Scholar |
Vajta, G, Rienzi, L, and Ubaldi, FM (2015). Open versus closed systems for vitrification of human oocytes and embryos. Reproductive BioMedicine Online 30, 325–333.
| Open versus closed systems for vitrification of human oocytes and embryos.Crossref | GoogleScholarGoogle Scholar |
Vanhecke, D, Zuber, B, Brugger, SD, and Studer, D (2012). Safe high-pressure freezing of infectious micro-organisms. Journal of Microscopy 246, 124–128.
| Safe high-pressure freezing of infectious micro-organisms.Crossref | GoogleScholarGoogle Scholar |
Zhang, F, Zhang, Z-Y, Cai, M-D, Li, X-X, Li, Y-H, Lei, Y, and Yu, X-L (2020). Effect of vitrification temperature and cryoprotectant concentrations on the mRNA transcriptome of bovine mature oocytes after vitrifying at immature stage. Theriogenology 148, 225–235.
| Effect of vitrification temperature and cryoprotectant concentrations on the mRNA transcriptome of bovine mature oocytes after vitrifying at immature stage.Crossref | GoogleScholarGoogle Scholar |
Zheng, X, Guo, W, Zeng, L, Zheng, D, Yang, S, Xu, Y, Wang, L, Wang, R, Mol, BW, Li, R, and Qiao, J (2022). In vitro maturation without gonadotropins versus in vitro fertilization with hyperstimulation in women with polycystic ovary syndrome: a non-inferiority randomized controlled trial. Human Reproduction 37, 242–253.
| In vitro maturation without gonadotropins versus in vitro fertilization with hyperstimulation in women with polycystic ovary syndrome: a non-inferiority randomized controlled trial.Crossref | GoogleScholarGoogle Scholar |
