Effect of different cryopreservation protocols on the metaphase II spindle in human oocytes.

The aim of this study was to evaluate the impact of different cryopreservation protocols on the repolymerization of metaphase (M)II spindles in human oocytes. Fresh aspirated donor oocytes were cryopreserved 3-4 h after retrieval using four different protocols: slow freezing using 1.5 mol/l 1,2-propanediol (PROH) + 0.2 mol/l sucrose (n = 36); 1.5 mol/l PROH + 0.3 mol/l sucrose (n = 34); 1.5 mol/l PROH + 0.3 mol/l sucrose with Na(+) depleted-choline replaced media (n = 27), and vitrification by the Cryotip method (n = 23). The control group comprised 34 fresh oocytes. Three hours after thawing, surviving and control oocytes were fixed for meiotic spindle/chromatin assessment. Survival rates were 63.8, 73.5, 74.1 and 86.9% respectively for the four protocols described above. Survival for vitrified oocytes was higher than that observed for slow freezing with 0.2 and 0.3 mol/l sucrose (P < 0.05). The proportion of oocytes showing normal spindle configuration was similar for the four protocols (81, 73.9, 88.9 and 81.3% respectively) and 88.5% for controls, showing that the MII spindle returns to its normal configuration after 3 h of post-thawing incubation under standard conditions, irrespective of the cryopreservation technique used.

Vitrification of preimplantation genetically diagnosed human blastocysts and its contribution to the cumulative ongoing pregnancy rate per cycle by using a closed device.

OBJECTIVE: To evaluate the survival rate and clinical results of our vitrification procedure on preimplantation genetic diagnosis (PGD) blastocysts and to calculate its actual contribution to the reproductive outcome per cycle. DESIGN: Retrospective clinical study. SETTING: University Institute IVI, Valencia, Spain. PATIENT(S): Patients who requested cryotransfer of surplus PGD blastocysts after failed fresh elective transfer. INTERVENTION(S): Retrospectively collected data during 2 years of experience with blastocyst vitrification. MAIN OUTCOME MEASURE(S): Primary outcome measures were the following: blastocyst recovery and survival; cryotransfer cancellation; and the implantation, pregnancy (PR), and ongoing-pregnancy rates. The secondary outcome measure was cumulative ongoing PR (COPR). RESULT(S): Cocultured vitrified PGD blastocysts were recovered and progressed in development after overnight culture (survival rate) at rates comparable to those of non-PGD blastocysts (49% and 42%, respectively). After transfer to 64% of patients, no statistical differences were found between PGD and non-PGD blastocyst groups concerning the following: PR (44% vs. 37%), implantation rate (40% vs. 27%), and ongoing-pregnancy rate (32% vs. 37%). Moreover, blastocyst vitrification significantly increased the COPR in both PGD and non-PGD cycles, from 47% (62/133) to 53% (70/133) and from 45% (24/53) to 53% (28/53), respectively. CONCLUSION(S): A preimplantation genetic diagnosis blastocyst vitrification procedure showed survival rates and improvements on the COPR that were comparable to those in non-PGD blastocyst cycles. Moreover, vitrification of biopsied and diagnosed embryos at the more advanced stages instead of at earlier cleavage stages is presented as an attractive strategy to consider in PGD programs.