Abstract
Refined slow freezing and new vitrification methods have significantly improved cryopreservation of human oocytes and embryos. Slow freezing techniques using a programmed cryo-machine have been traditionally employed, but usually take several hours to complete and involve ice-crystal formation during freezing. Vitrification has become a reliable strategy, not only because it is very simple but also because it can lead to high survival rates and viability. The underlying concept of vitrification is to transform the cells into an amorphous glassy state inside and outside the vitrified cell, instead of ice crystal formation. This requires high concentrations of cryoprotectancts (CPAs) and rapid cooling and warming steps. Conventional vitrification, using cryostraws with relatively high concentration of CPAs was not clinically effective compared with slow freezing, especially in oocyte and blastocyst. However, ultra-rapid vitrification, using a minimized volume of vitrification solution, has been adopted and could dramatically improve survival rate and viability. In order to vitrify the cells in liquid nitrogen, sufficient dehydration and permeation of the CPA are necessary, in any stage of gametes and embryos, although this ultra-rapid vitrification does not require full equilibration of CPAs because of its extremely high cooling rate. This is why this approach could decrease the concentration of CPAs to reach the vitrified state during the cooling and warming steps that could reduce the toxicity of CPA without forming ice-crystals inside the cell. Recently cryoloop, cryotop, hemistraw, open pulled straw (OPS), and EM grid were introduced as tools to minimize the vitrification solution at the final steps. In this chapter, the basic concepts and methodologies of both conventional vitrification and ultra-rapid vitrification will be discussed.
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Mukaida, T. (2009). The Use of Oocyte and Embryo Vitrification in Assisted Reproductive Technology. In: Voorhis, B., Schlegel, P., Racowsky, C., Carrell, D. (eds) Biennial Review of Infertility. Humana Press. https://doi.org/10.1007/978-1-60327-392-3_15
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DOI: https://doi.org/10.1007/978-1-60327-392-3_15
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