Cryopreservation is one of the keystones in clinical infertility treatment. Especially vitrification has become a well-established and widely used routine procedure that allows important expansion of therapeutic strategies when in vitro fertilization (IVF) is used to treat infertility. Vitrification of human blastocysts allows us to maximize the potential for conception from any one in vitro fertilization cycle and prevents wastage of embryos. This goes even further toward to best utilize a patient’s supernumerary oocytes after retrieval, maximizing the use of embryos from a single stimulation cycle. The technology may even be used to eliminate fresh embryo transfers for reasons of convenience, uterine receptivity, fertility preservation, preimplantation genetic diagnosis, or emergency management. In this chapter, the application of vitrification technology for cryopreserving human blastocyst will be revealed through step-by-step protocols. The results that are presented using the described protocols underscore the robustness of the vitrification technology for embryo cryopreservation.
Human blastocyst Cryopreservation Embryo transfer Cryostorage Vitrification
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
The author wants to thank the Fertility Centers of Illinois (FCI) and the embryologists at the FCI IVF Laboratory River North (Elissa Pelts, BS; Jill Matthews, BS; Sara Sanchez, BS; Rebecca Brohammer, BS; Yuri Wagner, BS; and Ewelina Pawlowska, MS) for their invaluable contributions and support in pushing vitrification to become our standard protocol for cryopreservation of human oocytes and blastocysts within our program since 2004.
Fahy GM, MacFarlane DR, Angell CA, Meryman HT (1984) Vitrification as an approach to cryopreservation. Cryobiology 21:407–426CrossRefGoogle Scholar
Fahy GM (1986) Vitrification: a new approach to organ cryopreservation. In: Merryman HT (ed) Transplantation: approaches to graft rejection. Alan R Liss, New York, pp 305–335Google Scholar
Seki S, Mazur P (2009) The dominance of warming rate over cooling rate in the survival of mouse oocytes subjected to a vitrification procedure. Cryobiology 59:75–82CrossRefGoogle Scholar
Mazur P, Seki S (2011) Survival of mouse oocytes after being cooled in a vitrification solution to−196 °C at 95 to 70,000 °C/min and warmed at 610° to 118,000 °C/min: A new paradigm for cryopreservation by vitrification. Cryobiology 62:1–7CrossRefGoogle Scholar
Rall WF, Fahy GM (1985) Ice-free cryopreservation of mouse embryos at−196 °C by vitrification. Nature 313:573–575CrossRefGoogle Scholar
Ali J, Shelton JN (1993) Vitrification of preimplantation stages of mouse embryos. J Reprod Fertil 98:459–465CrossRefGoogle Scholar
Vajta G, Holm P, Kuwayama M, Booth PJ, Jacobsen H, Greve T, Callesen H (1998) Open pulled straws (OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod Dev 51:53–58CrossRefGoogle Scholar
Kuleshova L, Gianaroli L, Magli C, Ferraretti A, Trounson A (1999) Birth following vitrification of a small number of human oocytes: case report. Hum Reprod 14:3077–3079CrossRefGoogle Scholar
Yoon TK, Chung HM, Lim JM, Han SY, Ko JJ, Cha KY (2000) Pregnancy and delivery of healthy infants developed from vitrified oocytes in a stimulated in vitro fertilization-embryo transfer program. Fertil Steril 74:180–181CrossRefGoogle Scholar
Walker DL, Tummon IS, Hammitt DG, Session DR, Dumesic DA, Thornhill AR (2004) Vitrification versus programmable rate freezing of late stage murine embryos: a randomized comparison prior to application in clinical IVF. Reprod Biomed Online 8:558–568CrossRefGoogle Scholar
Takahashi K, Mukaida T, Goto T, Oka C (2005) Perinatal outcome of blastocyst transfer with vitrification using cryoloop: a 4-year follow-up study. Fertil Steril 84:88–92CrossRefGoogle Scholar
Liebermann J, Tucker MJ (2006) Comparison of vitrification versus conventional cryopreservation of day 5 and day 6 blastocysts during clinical application. Fertil Steril 86:20–26CrossRefGoogle Scholar
Liebermann J (2009) Vitrification of human blastocysts: an update. Reprod Biomed Online 19(Suppl 4):105–114CrossRefGoogle Scholar
Liebermann J (2011) More than six years of blastocyst vitrification—what is the verdict? US Obstet Gynecol 5:14–17Google Scholar
Liebermann J, Tucker MJ (2002) Effect of carrier system on the yield of human oocytes and embryos as assessed by survival and developmental potential after vitrification. Reproduction 124:483–489CrossRefGoogle Scholar
Liebermann J, Nawroth F, Isachenko V, Isachenko E, Rahimi G, Tucker MJ (2002) Potential importance of vitrification in reproductive medicine. Biol Reprod 67:1671–1680CrossRefGoogle Scholar
Liebermann J, Dietl J, Vanderzwalmen P, Tucker MJ (2003) Recent developments in human oocyte, embryo and blastocyst vitrification: where are we now? Reprod Biomed Online 7:623–633CrossRefGoogle Scholar