Cryopreservation of Mammalian Gametes and Embryos

Matthew VerMilyea, Amber Brewer, VerMilyea, Matthew, Brewer, Amber

This chapter will describe the use of the Irvine Scientific(®) vitrification and warming solutions (Vit Kits(®)) along with a detailed protocol for the correct use of the CryoTip(®) vitrification device for human embryos and oocytes. Successful pregnancies have been reported after carrying out rapid vitrification methods of oocytes, cleavage stage embryos, and blastocysts using the CryoTip(®) (Popwell et al. Fertil Steril, 101:e20, 2014; Kuwayama et al. Reprod Biomed 11:608-614, 2005; Kuwayama et al. Fertil Steril 84:S187, 2005; Kuwayama et al. Vitrification of human embryos using the CryoTip TM method. Reprod Biomed 11:608-614). Compared to other vitrification carrier devices, the CryoTip(®) is considered a closed carrier for vitrification, thereby eliminating the theoretical risk of disease transmission through contaminated liquid nitrogen during cooling and storage (Bielanski et al. Cryobiology, 40:110-116, 2000). The CryoTip(®) is cleared by the FDA and has CE mark approval as a closed device to carry gametes or embryos in a specialized medium during cryopreservation procedures and subsequent long-term storage in liquid nitrogen. The CryoTip(®) has been shown to be a safe and reliable vitrification device, and when compared to other open system vitrification devices, it has provided similar results (Martino et al. Reprod Biol Endocrinol 11:27, 2013, Valbuena et al. Fertil Steril 97:209-217, 2012; Kuwayama et al. Reprod Biomed 11:608-614, 2005). The CryoTip(®) has also been shown to be suitable for use as a vitrification device for the cryopreservation of small volumes of sperm (Tanaka et al. Fertil Steril 90: S292, 2008).