Details on the findings of standard freezing and thawing protocols for cryopreservation of biopsied human embryos at the blastocyst stage

                                                                                         

The combination of in vitro fertilization (IVF) and pre-implantation genetic diagnosis (PGD) for genetic studies and aneuploid screening has become common in recent years. PGD has been used for known single-gene disorders (1), repeated miscarriage (2), advanced maternal age (3), X-linked diseases (4), Translocations (5),repetitive IVF failure (6), severe male infertility (7), and routine aneuploid screening (8).

It gives the couples an option to transfer only unaffected embryos. Therefore, the pregnancy is initiated with the knowledge that the fetus is free from diseases. Biopsy protocols include blastomere, polar body and trophoblast biopsy. The most commonly employed is cleavage stage biopsy performed at 6-10 cell stage. Usually one or two blastomeres are removed (9-10). The opening created in the zona pellucida during the biopsy may reduce the subsequent embryos susceptible to cryopreservation damage. Finding a safe cryopreservation method for biopsied human embryos is paramount to allow for future use of remaining normal embryos.

Further more, certain single-gene disorder diagnosis is protracted, requesting freezing of all biopsied embryos (11). So far, there is insufficient data on the optimal freezing method for biopsied human embryos .There are several reports of modified cryopreservation methods for biopsied human embryos (12-16). In this study, we used standard freezing and thawing procedures for blastocyst to freeze and thaw biopsied human embryos at blastocyst stage by comparing embryo survival and further development, as well as implantation and pregnancy rates between biopsied and non-biopsied embryos in order to test weather the standard protocol of blastocyst cryopreservation is suitable for biopsied blastocyst.

To the best of our knowledge, this is the first report to use standard blastocyst freezing and thawing protocols for biopsied embryo cryopreservation.

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