This paper reports the use of a novel automatic vitrification device (Sarah, Fertilesafe, Israel) for cryopreservation of oocytes and embryos.
Mice oocytes (n = 40) and embryos (8 cells, n = 35 and blastocysts, n = 165), bovine embryos (2PN, n = 35), and MII oocytes (n = 84) were vitrified using this automated device. A total of 42 (2 cells) mice embryos, 20 (2PN) bovine embryos, and 150 MII bovine oocytes were used as fresh controls and grown to blastocysts. Upon rewarming, all were assessed for viability, cleavage, blastocyst, and hatching rates.
Ninety-five % (38/40) of the mice MII oocytes regained isotonic volumes and all (100%) the surviving were viable. Rewarmed 8-cell mice embryos had 95% (33/35) blastulation rate and 80% (28/35) hatched. Rewarmed mice blastocysts had 97% survival rate (160/165) and 81% (135/165) hatched. Fresh control mice embryos had 100% (42/42) blastulation and 73% (21/42) hatching rates. Bovine embryos’ survival was 100% with 54% (19/35) cleavage and 9% (3/35) blastulation rate. Fresh control bovine embryos had 65% (13/20) cleavage and 20% (4/20) blastulation rate. Vitrified bovine oocytes had 100% survival (84/84), 73% (61/84) cleavage, and 7% (6/84) blastocysts’ rates; fresh control had 83% (125/150) cleavage and 11% (17/150) blastocysts’ rates.
This novel automatic vitrification device is capable to produce high survival rates of oocytes and embryos. We anticipate that as the demand for vitrification of gametes, embryos, and reproductive tissues increases worldwide, the availability of an automated vitrification device will become indispensable for standardization, simplification, and reproducibility of the entire process.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
American Society for Reproductive Medicine. Mature oocyte cryopreservation: a guideline. Practice committees of American Society for Reproductive Medicine, Society for Assisted Reproductive Technology. Fertil Steril. 2013;99:37–43.
Cobo A, Garcia-Velasco JA, Domingo J, Remohi J, Pellicer A. Is vitrification of oocytes useful for fertility preservation for age-related fertility decline and in cancer patients? Fertil Steril. 2013;99:1485–95.
Rudick B, Opper N, Paulson R, Bendikson K, Chung K. The status of oocyte cryopreservation in the United States. Fertil Steril. 2010;94:2642–6.
Rienzi L, Gracia C, Maggiulli R, LaBarbera AR, Kaser DJ, Ubaldi FM, et al. Oocyte, embryo and blastocyst cryopreservation in ART: systematic review and meta-analysis comparing slow freezing versus vitrification to produce evidence for the development of global guidance. Hum Reprod Update. 2017;23(2):139–55.
Arav A. Cryopreservation of oocytes and embryos. Theriogenology. 2014;81(1):96–102.
Mazur P, Schneider U, Mahowald AP. Characteristics and kinetics of subzero chilling injury in Drosophila embryos. Cryobiology. 1992;29(1):39–68.
Arav A. Cryopreservation of oocytes and embryos, DVM thesis 1989.
Saragusty J, Arav A. Current progress in oocyte and embryo cryopreservation by slow freezing and vitrification. Reproduction. 2011;141(1):1–19.
Rall WF, Fahy GM. Ice-free cryopreservation of mouse embryos at -196 degrees C by vitrification. Nature. 1985;313(6003):573–5.
Arav A. Vitrification of oocytes and embryos. In: Lauria A, Gandolfi F, editors. Embryonic development and manipulation in animal reproduction. Portland press; 1992: 255–264.
Kuwayama M. Highly efficient vitrification for cryopreservation of human oocytes and embryos: the Cryotop method. Theriogenology. 2007;67(1):73–80.
Levi-Setti PE, Patrizio P, Scaravelli G. Evolution of human oocyte cryopreservation: slow freezing versus vitrification. Curr Opin Endocrinol Diabetes Obes. 2016;23(6):445–50.
Roy TK, Brandi S, Tappe NM, Bradley CK, Vom E, Henderson C, et al. Embryo vitrification using a novel semi-automated closed system yields in vitro outcomes equivalent to the manual Cryotop method. Hum Reprod. 2014;29(11):2431–8.
Alpha Scientists in Reproductive Medicine. The alpha consensus meeting on cryopreservation key performance indicators and benchmarks: proceedings of an expert meeting. Reprod BioMed Online. 2012;25:146–67.
Kader AA, Choi A, Orief Y, Agarwal A. Factors affecting the outcome of human blastocyst vitrification. Reprod Biol Endocrinol. 2009;7:99.
Kalo D, Roth Z. Effects of mono(2-ethylhexyl)phthalate on cytoplasmic maturation of oocytes—the bovine model. Reprod Toxicol. 2015;53:141–51.
Arav A, Natan Y, Levi-Setti PE, Menduni F, Patrizio P. New methods for cooling and storing oocytes and embryos in a clean environment of −196°C. Reprod BioMed Online. 2016;33(1):71–8.
Cobo A, García-Velasco JA. Why all women should freeze their eggs. Curr Opin Obstet Gynecol. 2016;28(3):206–10.
Cobo A, García-Velasco JA, Coello A, Domingo J, Pellicer A, Remohí J. Oocyte vitrification as an efficient option for elective fertility preservation. Fertil Steril. 2016;105(3):755–764.e8.
Cobo A, Garrido N, Pellicer A, Remohí J. Six years’ experience in ovum donation using vitrified oocytes: report of cumulative outcomes, impact of storage time, and development of a predictive model for oocyte survival rate. Fertil Steril. 2015;104(6):1426–34.
Nagy ZP, Nel-Themaat L, Chang CC, Shapiro DB, Berna DP. Cryopreservation of eggs. Methods Mol Biol. 2014;1154:439–54.
Cobo A, Vajta G, Remohí J. Vitrification of human mature oocytes in clinical practice. Reprod BioMed Online. 2009;19(Suppl 4):4385.
Nagy ZP, Chang CC, Shapiro DB, Bernal DP, Kort HI, Vajta G. The efficacy and safety of human oocyte vitrification. Semin Reprod Med. 2009;27(6):450–5.
Liebermann J. Chapter 11 human embryo Vitrification. Methods Mol Biol. 2017;1568:141–59.
Fasano G, Fontenelle N, Vannin AS, Biramane J, Devreker F, Englert Y, et al. A randomized controlled trial comparing two vitrification methods versus slow-freezing for cryopreservation of human cleavage stage embryos. J Assist Reprod Genet. 2014;31(2):241–7.
Loutradi KE, Kolibianakis EM, Venetis CA, Papanikolaou EG, Pados G, Bontis I, et al. Cryopreservation of human embryos by vitrification or slow freezing: a systematic review and meta-analysis. Fertil Steril. 2008;90(1):186–93.
Shi Q, Xie Y, Wang Y, Li S. Vitrification versus slow freezing for human ovarian tissue cryopreservation: a systematic review and meta-anlaysis. Sci Rep. 2017;7(1):8538.
Silber S. Ovarian tissue cryopreservation and transplantation: scientific implications. J Assist Reprod Genet. 2016;33(12):1595–603.
Silber S. Chapter 13 human ovarian tissue vitrification. Methods Mol Biol. 2017;1568:177–94.
Arav A, Shehu D, Mattioli M. Osmotic and cytotoxic study of vitrification of immature bovine oocyte. J Reprod Fert. 1993;99:353–8.
Ghetler Y, Yavin S, Shalgi R, Arav A. The effect of chilling on membrane lipid phase transition in human oocytes and zygotes. Hum Reprod. 2005;20(12):3385–9.
Zenzes MT, Bielecki R, Casper RF, Leibo SP. Effects of chilling to 0 degrees C on the morphology of meiotic spindles in human metaphase II oocytes. Fertil Steril. 2001;75(4):769–77.
Yavin S, Arav A. Measurement of essential physical properties of vitrification solutions. Theriogenology. 2007;67(1):81–9.
Martino A, Pollard JW, Leibo SP. Effect of chilling bovine oocytes on their developmental competence. Mol Reprod Dev. 1996;45(4):503–12.
Morató R, Izquierdo D, Paramio MT, Mogas T. Cryotops versus open-pulled straws (OPS) as carriers for the cryopreservation of bovine oocytes: effects on spindle and chromosome configuration and embryo development. Cryobiology. 2008;57(2):137–41.
Do VH, Walton S, Catt S, Taylor-Robinson AW. A comparative analysis of the efficacy of three cryopreservation protocols on the survival of in vitro-derived cattle embryos at pronuclear and blastocyst stages. Cryobiology. 2017;77:58–63.
Massip A, Van der Zwalmen P, Leroy F. Effect of stage of development on survival of mouse embryos frozen–thawed rapidly. Cryobiology. 1984;21:574–7.
Camus M, Van Den Abbeel E, Van Waesberghe L, Wisanto A, Devroey P, Van Steirteghem A. Human embryo viability after freezing with dimethylsulfoxide as a cryoprotectant. Fertil Steril. 1989;51(3):460–5.
About this article
Cite this article
Arav, A., Natan, Y., Kalo, D. et al. A new, simple, automatic vitrification device: preliminary results with murine and bovine oocytes and embryos. J Assist Reprod Genet 35, 1161–1168 (2018). https://doi.org/10.1007/s10815-018-1210-9