Improved outcome of frozen-thawed blastocyst transfer with Menezo's two-step thawing compared to the stepwise thawing protocol

Purpose: To compare the effects of two different blastocyst thawing protocols on implantation, pregnancy, and live birth rates. Methods: Ninety four consecutive frozen-thawed blastocyst transfers from 1996 to 2002 were retrospectively analyzed. Blastocysts were cryopreserved using Menezo's two-step slow-freezing protocol. Frozen blastocysts were thawed for transfer according to Menezo's stepwise or two-step protocol. Immediately after thawing, assisted hatching was performed and all embryos were cultured for 3 to 5 h before transfer. Only viable embryos were selected for transfer. Results: Implantation, pregnancy ( determined by the presence of fetal cardiac activity), and live birth rates were significantly higher with two-step ( 25, 45.7, and 42.9%) than with stepwise thawing protocol (9.2, 18.6, and 16.9%, P < 0.01). The percentage of gestational sacs resulting in live babies was higher with two-step thawing ( 76.6%) than with stepwise thawing (50%, P = 0.04). Conclusions: We conclude that the two-step thawing protocol offers advantages over the stepwise method. The two-step thawing protocol dramatically increased embryo implantation potential, resulting in higher pregnancy rate, and subsequent live birth rate, after frozen blastocyst transfer.