Abstract

Vitrification is a cryo-preservation method often used in embryos obtained from mares or jennies. It consists in the dramatic reduction of temperature to levels close to -196 °C, that allows the cryopreserving solution containing the embryo to pass from liquid to vitreous state. Several improvements to vitrification protocols have made possible to cryo-preserve embryos with different sizes; since during the first decade after the year 2000, only small embryos were successfully vitrified. Embryos collected at the sixth day post ovulation (PO) are usually smaller or equal to 300 micrometers in diameter (≤ 300 µmØ) and can be routinely vitrified following simple protocols; they have a higher post vitrification pregnancy rate (PVPR) when compared to large embryos which have more than 300 micrometers in diameter (˃ 300 µmØ). The high PVPR of embryos ≤ 300 µmØ is due to an embryo capsule (EC) that is not fully developed yet and has a high permeability to cryo-preserving solutions. At present time, embryos collected either the seventh or eighth day PO are ˃ 300 µmØ and are characterized to have a low post vitrification survival; in order to increase their PVPR their EC might be punctured to make it permeable to cryopreserving solutions. Additionally, there are at least two factors that can be manipulated to increase the PVPR of embryos ˃ 300 µmØ; one is to reduce their size by aspiring their blastocoelic liquid (BL), and the other is to induce a high temperature transfer index (TTI) to rapidly reach -196 °C.

Full Text
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