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Vitrification is an exciting new technique which is used to cryopreserve (freeze) supernumerary eggs and embryos. Using ultrarapid (flash) freezing techniques, it's now possible to efficiently store spare eggs and embryos, so they can be used later on to achieve a pregnancy after thawing. Survival rates after vitrification are better than they were with conventional slow freezing.
This describes the laboratory protocol we use for freezing eggs and embryos.
The principle is simple. Vitrification is dependent on the placement of the oocyte/embryo in a very small volume of vitrification medium which is then cooled at an extreme rate, so that the embryos are flash frozen and preserved intact.
Media : CVM Vitrification Medium (Supplier :COOK IVF)
The vitrification kit (K-SIBV-5000) is a three-step freezing system containing Cryobase buffer with ethylene glycol, DMSO and trehalose as the cryoprotectants.The kit has 4 vials.
Preparation of Vitrification Solution 2 and Vitrification Solution 3
Remove solution 4 from refrigerator and keep it for thawing , as solution 4 contains DMSO, it always becomes ice once refrigerated.
Vitrification Solution 2 = 400ul Solution 4 + 4.6 ml Solution 2
Vitrification Solution 3 = 1 ml Solution 4 + 5.25 ml of Solution 3
Note : loading embryos onto cryoloop is the most difficult part in the procedure. One has to make sure that the volume of media should be as low as possible, since the freezing medium a has high concentration of cryoprotectant and is embryotoxic, which can affect the survival of embryos.