AMH levels have been used as reliable indicators for ovarian response to gonadotropins, and it has been shown to be closely associated with oocyte yield and to some extent with clinical outcomes in human IVF treatment cycles. However, it remains controversial whether AMH is associated not only with oocyte quantity but possibly also with oocyte quality. To compare cycle outcomes from cycles with variable oocyte numbers, we looked at recipient cycles using cryo-banked warmed oocytes from two groups of oocyte donors; those with AMH levels below 5ng/ml and those with an AMH greater than 5 where 5 represented the median AMH value for the data set. Once segregated around the median AMH value, we compared outcomes for multiple parameters as a function of AMH. Retrospective study. Oocyte donors recruited for our donor egg bank were required to have an AMH of at least 2.0ng/ml and a basal antral follicle count of at least 20 (donors with PCOS and with other conditions not fulfilling the established requirements were excluded). All donors were treated with rFSH/GnRH antagonist and using GnRH agonist as trigger, with transvaginal egg collection occurring 36 hours after the trigger injection. Cryopreservation of donor oocytes derived from 274 oocyte retrievals was performed using minimum volume vitrification. Oocytes were warmed for each matched recipient independently (402 recipient cycles, total of 2707 oocytes warmed). Outcome data was grouped by serum AMH level; I) <5 ng/ml, II) ≥5 ng/ml. Outcome parameters were analyzed by Chi-square. There were no cases of OHSS among the donors. All other variables, including semen parameters were similar in both groups. Tabled 1AMH<5AMH>=5p valueTotal # of retrieval cycle for donation138136-AMH (mean±SD)(ng/ml)3.37±2.947.86±3.27p<0.05# of retrieved follicle (mean±SD)31.74±4.9540.89±15.67p<0.05Total # of recipient cycle200202-Total # of oocyte warmed (mean±SD)1327(6.64±1.70)1380(6.83±1.75)NSTotal # of oocyte survived (%)1247(93.97)1295(93.84)NSTotal # of oocyte fertilized (%)1029(77.50)1032(74.78)NSTotal # of blastocyst (d5/6) (%)744(56.06)697(50.50)p<0.05Total # of ET (mean±SD)222(1.11±0.31)229(1.13±0.34)NSTotal # of implantation (%)134(60.36)122(53.27)NSTotal # of clinical pregnancy (%)122(61.00)108(53.46)NS Open table in a new tab Our results demonstrate that oocyte yield is strongly correlating with AMH levels in oocyte donors. Survival and fertilization rates after warming were not significantly different between low and high AMH groups. Similarly, pregnancy and implantation rates were not significantly different between groups. It is noteworthy that the blastocyst conversion rate was slightly (but statistically significant) lower when the donors' AMH level was equal to and/or above 5 ng/ml. Our findings imply that AMH can be a good indicator for oocyte yield in oocyte donors. All laboratory and clinical outcomes were similar between low and high AMH groups except for the blastulation rate, which warrants further study.

Long distance/cross border egg donation is driven largely by restrictive law, cost, and accessibility of egg donors. Current egg donation methods require that either intended parents travel to clinics where donor eggs are located, or the vitrified donor eggs must be shipped to clinics where intended parents will do IVF and ET. Using a cryo-donor egg bank, donor eggs can be shipped to receiving clinics for long distance/cross border egg donation cases. However, it is not uncommon for there to be no high quality embryos available after a donor egg warming cycle. In order to avoid this frustrating and disappointing situation, we developed a novel model for long distance/cross border egg donation, with the objective being to test whether this model can provide the benefit of remaining at home for intended parents while simultaneously guaranteeing high quality embryo creation. Retrospective study. All donors were submitted to COHS using GnRH antagonist with rFSH and GnRH agonist trigger. Cryopreservation of donor oocytes was performed using minimum volume vitrification. A total of 484 donor oocytes (54 individual donors) from the donor oocyte cryo-bank were matched to 70 recipients (22 recipient clinics) and were included in this study. The frozen sperm specimen of intended parents was delivered from remote clinics for the donor egg warming and insemination. ICSI using thawed sperm was performed after 3 hours of in vitro culture and osmotic equilibration. The fertilized eggs were subsequently cultured to day-5/day-6 and embryos were vitrified at blastocyst stage. Cryopreserved embryos were then shipped to the recipient location for warming and transfer. Tabled 1No. of recipient cycle70No. of egg warmed (mean±sd)484 (6.91±2.10)No. of egg survived / fertilized (% per egg warmed)453 (93.5) / 395 (81.6)No. of day 5 / day 6 blastocyst (% per egg warmed)144 (29.7) / 79 (16.3)No. of blastocyst vitrified (mean±sd)223 (3.18±1.48)No. of Frozen ET cycle(cumulative)88No. of Embryo Transferred (mean±sd)122 (1.38±0.49)No. of Clinical Pregnancy (cumulative) (%)44 (62.8)No. of implantation (%)53 (43.4)No. of recipient delivered25No. of live birth29No. of ongoing pregnancy18 Open table in a new tab The study demonstrates that long distance/cross border egg donation cycles can be operated efficiently by using the combination of the transport of frozen husband/partner sperm to the donor egg bank for fertilization followed by return shipment of resulting embryos to the recipient clinic. Since the embryos are created on-site at the donor egg bank, the quality and quantity of the resulting embryos can be assured before returning them to recipient clinic for subsequent embryo warming and transfer. Long distance/cross border egg donation can be operated by this model without increase in tangible and intangible costs relative to care received in one's own community. By eliminating the need to travel, this model of embryo creation allows intended parents to get a minimum number of high quality embryos guaranteed while concurrently preventing dislocation from local support networks.

ObjectiveImmature oocytes from superovulated cycles can undergo the final stages of meiotic maturation spontaneously in vitro, especially, from metaphase I (MI) stage. In order to reach the optimal fertilization time window for ICSI, oocyte vitrification in combination with in-vitro culture might offer a solution to provide additional usable eggs. Therefore, the objective was to evaluate the efficiency of MI-II vs. MII sibling oocytes after oocyte vitrification.DesignProspective study.Materials and MethodsA total of 188 donor oocytes (139 MII and 49 MI-MII) used from the donor oocyte cryo-bank for 25 matched recipients were included. Donor sibling oocytes were divided into two groups for vitrification: in-vivo matured oocytes (MII, A) vs. MI in-vitro matured to MII oocytes (MI-II, B). The incubated MI oocytes were evaluated for maturation after 3 hours of removal of cumulus cell complex, and then those which extruded their PB (MI-II oocytes, B) were vitrified. Sibling oocytes from both groups were warmed at the same time. Oocytes were fertilized by ICSI after 3 hours of in vitro culture post warming. On day 5, blastocyst formation was assessed and embryo transfer was performed. Results were analyzed by the Chi-square (P<0.05) statistical test.ResultsTabled 1A (MII)B(MI-II)Recipient age (N=25)39.8 ± 5.1Donor age (N=21)26.7 ± 2.6# of oocyte warmed139 (5.5±1.2)49 (1.9±0.8)# of oocyte survived122/139 (87.7)43/49 /(87.7)# of oocyte fertilized114/139 (82.0)35/49 (71.4)# of day5/6 blastocyst*Significantly different (P<0.05).75/139 (53.9)9/49 (18.3)# of embryo selected for transfer*Significantly different (P<0.05).40/139 (28.7)4/49 (8.1)# of ET44 (1.76 ± 0.59)Clinical pregnancy rate20/25 (80.0)Implantation rate26/44 (59.0)* Significantly different (P<0.05). Open table in a new tab ConclusionThe oocyte survival and fertilization rates were comparable between two groups.The use of oocyte vitrification on in vitro matured MI-II oocytes may be worthwhile not only to increase the number of injectable oocytes at the time of ICSI, but also it could preserve some of best quality oocytes for patients. ObjectiveImmature oocytes from superovulated cycles can undergo the final stages of meiotic maturation spontaneously in vitro, especially, from metaphase I (MI) stage. In order to reach the optimal fertilization time window for ICSI, oocyte vitrification in combination with in-vitro culture might offer a solution to provide additional usable eggs. Therefore, the objective was to evaluate the efficiency of MI-II vs. MII sibling oocytes after oocyte vitrification. Immature oocytes from superovulated cycles can undergo the final stages of meiotic maturation spontaneously in vitro, especially, from metaphase I (MI) stage. In order to reach the optimal fertilization time window for ICSI, oocyte vitrification in combination with in-vitro culture might offer a solution to provide additional usable eggs. Therefore, the objective was to evaluate the efficiency of MI-II vs. MII sibling oocytes after oocyte vitrification. DesignProspective study. Prospective study. Materials and MethodsA total of 188 donor oocytes (139 MII and 49 MI-MII) used from the donor oocyte cryo-bank for 25 matched recipients were included. Donor sibling oocytes were divided into two groups for vitrification: in-vivo matured oocytes (MII, A) vs. MI in-vitro matured to MII oocytes (MI-II, B). The incubated MI oocytes were evaluated for maturation after 3 hours of removal of cumulus cell complex, and then those which extruded their PB (MI-II oocytes, B) were vitrified. Sibling oocytes from both groups were warmed at the same time. Oocytes were fertilized by ICSI after 3 hours of in vitro culture post warming. On day 5, blastocyst formation was assessed and embryo transfer was performed. Results were analyzed by the Chi-square (P<0.05) statistical test. A total of 188 donor oocytes (139 MII and 49 MI-MII) used from the donor oocyte cryo-bank for 25 matched recipients were included. Donor sibling oocytes were divided into two groups for vitrification: in-vivo matured oocytes (MII, A) vs. MI in-vitro matured to MII oocytes (MI-II, B). The incubated MI oocytes were evaluated for maturation after 3 hours of removal of cumulus cell complex, and then those which extruded their PB (MI-II oocytes, B) were vitrified. Sibling oocytes from both groups were warmed at the same time. Oocytes were fertilized by ICSI after 3 hours of in vitro culture post warming. On day 5, blastocyst formation was assessed and embryo transfer was performed. Results were analyzed by the Chi-square (P<0.05) statistical test. ResultsTabled 1A (MII)B(MI-II)Recipient age (N=25)39.8 ± 5.1Donor age (N=21)26.7 ± 2.6# of oocyte warmed139 (5.5±1.2)49 (1.9±0.8)# of oocyte survived122/139 (87.7)43/49 /(87.7)# of oocyte fertilized114/139 (82.0)35/49 (71.4)# of day5/6 blastocyst*Significantly different (P<0.05).75/139 (53.9)9/49 (18.3)# of embryo selected for transfer*Significantly different (P<0.05).40/139 (28.7)4/49 (8.1)# of ET44 (1.76 ± 0.59)Clinical pregnancy rate20/25 (80.0)Implantation rate26/44 (59.0)* Significantly different (P<0.05). Open table in a new tab ConclusionThe oocyte survival and fertilization rates were comparable between two groups.The use of oocyte vitrification on in vitro matured MI-II oocytes may be worthwhile not only to increase the number of injectable oocytes at the time of ICSI, but also it could preserve some of best quality oocytes for patients. The oocyte survival and fertilization rates were comparable between two groups.The use of oocyte vitrification on in vitro matured MI-II oocytes may be worthwhile not only to increase the number of injectable oocytes at the time of ICSI, but also it could preserve some of best quality oocytes for patients.

To study the blastocyst genetic constitution (possible chromosomal mosaicism between ICM and TE and within TE), after isolating ICM by a novel method.and to evaluate array-CGH accuracy against FISH testing Cross-sectional study. 32 blastocysts diagnosed as genetically abnormal by array-CGH were donated to research. ICM was isolated from the trophectoderm (TE) using a novel mechanical separation which was confirmed by KRT18 immunostaining. 24 of the 32 embryo had single or double aneuploidy and 8 aneusomic for unbalanced product of translocation or for long single arm deletions/duplications but with normal ploidy (by aCGH). After biopsy, individual samples were fixed by Carnoy method and analyzed with conventional 9-chromosomes FISH and locus specific probes. We reanalyzed 32 ICM and 96 TE samples (3 separate groups of cells from each TE of the same embryo). Looking at the single chromosome level, 28 out of 28 aneuploidies diagnosed by a-CGH were confirmed by FISH reanalysis both in ICM and TE samples resulting in a analytical sensitivity of 100%(95%CI 90.2-100). For almost all embryos, the reported aneuploidies were constitutive. As regard to chromosomes diagnosed as disomic, only 2 out of 248 (specificity 99.1 95%CI 97.4-99.8) were misdiagnosed by a-CGH, a trisomy 13 in an unbalanced embryo from reciprocal translocation carrier observed in a constitutive state and a mosaic nullisomy/monosomy 16 in a double aneuploid embryo. Consequently, only one out of 32 embryos was misdiagnosed as false negative. Only in one case we observed a confined trisomy 17 in all nuclei of a TE sample from a trisomic embryo for chromosome 13. In all other embryos ICM and TE samples were concordant and confirmed the original a-CGH diagnosis. Our data show a very high analytical accuracy of a-CGH to diagnose embryo chromosomal constitution. Concordance between ICM and TE was reported for all embryos analyzed, showing no sign of mosaicism or preferential allocation at blastocyst stage.

OBJECTIVE: Our objective was to compare oocyte vitrification outcomes with vitrification solution containing single permeable cryoprotectant (Ethylene glycol (EG)) vs. mixture of permeable cryoprotectants (EG and Dimethyl sulfoxide (DMSO)).DESIGN: Prospective study.MATERIALS AND METHODS: Cryopreservation on sibling oocytes from 28 donors was performed by minimum volume vitrification method. Donor sibling oocytes were divided into two groups for vitrification: mixtures of permeable cryoprotectants (15% EG and 15% DMSO; group A) vs. single permeable cryoprotectant (30% EG; group B). Sibling oocytes from both groups were warmed at the same time for each recipient (N=52). Oocytes were fertilized by ICSI 2-3 hours after warming. On day 5, blastocyst formation was assessed and embryo transfer was performed. Results were analyzed by Fisher's exact test (P<0.05).Tabled 1A (EG + DMSO)B (EG)Recipient age (N=52)41.0 ± 4.9Donor age (N=28)27.0 ± 2.9# of oocyte warmed216133# of oocyte survived (%)∗Significantly different (P<0.05).193/216 (89.3)128/133 (96.2)# of oocyte fertilized (%)175/216 (81.0)109/133 (81.9)# of blastocyst (%)124/216 (57.4)71/133 (53.4)# of embryo transferred (%)62/216 (28.7)41/133 (30.8)# of embryo re-vitrified (%)62/216 (28.7)30/133 (22.6)# of ET (mean ± SD)1.98 ± 0.3Clinical pregnancy rate (%)29 (55.8)Implantation rate (%)48/103 (46.5)# of delivery8# of live birth14∗ Significantly different (P<0.05). Open table in a new tab CONCLUSION: These data suggested that using single permeable cryoprotectant (EG) in the vitrification solutions had better oocyte survival than the mixtures of cryoprotectants (EG + DMSO). We also demonstrated that the fertilization and blastocyst rates were similar between single and mixtures of permeable cryoprotectant. The data presented here suggest a vitrification solution with only one permeable cryoprotectant can provide equivalent efficacy and efficiency of oocyte vitrification to a common protocol that contains cryoprotectant mixtures. OBJECTIVE: Our objective was to compare oocyte vitrification outcomes with vitrification solution containing single permeable cryoprotectant (Ethylene glycol (EG)) vs. mixture of permeable cryoprotectants (EG and Dimethyl sulfoxide (DMSO)). DESIGN: Prospective study. MATERIALS AND METHODS: Cryopreservation on sibling oocytes from 28 donors was performed by minimum volume vitrification method. Donor sibling oocytes were divided into two groups for vitrification: mixtures of permeable cryoprotectants (15% EG and 15% DMSO; group A) vs. single permeable cryoprotectant (30% EG; group B). Sibling oocytes from both groups were warmed at the same time for each recipient (N=52). Oocytes were fertilized by ICSI 2-3 hours after warming. On day 5, blastocyst formation was assessed and embryo transfer was performed. Results were analyzed by Fisher's exact test (P<0.05). CONCLUSION: These data suggested that using single permeable cryoprotectant (EG) in the vitrification solutions had better oocyte survival than the mixtures of cryoprotectants (EG + DMSO). We also demonstrated that the fertilization and blastocyst rates were similar between single and mixtures of permeable cryoprotectant. The data presented here suggest a vitrification solution with only one permeable cryoprotectant can provide equivalent efficacy and efficiency of oocyte vitrification to a common protocol that contains cryoprotectant mixtures.

OBJECTIVE: Our objective was to assess the efficacy and efficiency of oocyte donation cycles using oocyte cryo-banking in comparison to fresh oocyte donation cycles. DESIGN: Prospective, parallel-group study. MATERIALS AND METHODS: From Jan 2007 to Mar 2010, a total of 395 recipient cycles of donated oocytes were included in this study. Cryopreservation on oocytes was performed by minimum volume vitrification with 15% ethylene glycol, 15% DMSO, and 0.5M sucrose. Oocytes were fertilized by 40 hr after hCG administration or 2-3 hr after oocyte warming. Clinical pregnancy was confirmed by detecting fetal cardiac activity (FCA) with ultrasound. Results were analyzed by the One-way ANOVA or the Fisher's exact tests, as appropriate (P<0.05). RESULTS: As shown below.Tabled 1FreshVitrifiedP value# of Recipients148247NATotal # of donor cycles148139NARecipient age40.6 ± 4.741.0 ± 4.6NSDonor age26.7 ± 3.226.3 ± 2.7NS# of oocyte used/warmed (mean ± SD)3107 (20.9 ± 9.0)1592 (6.4 ± 2.0)<0.0001# of oocyte remained01187NA# of oocyte survived (%)-1386 (87.0)NA# of oocyte fertilized (mean ± SD)1823 (12.3 ± 5.3)1206 (4.8 ± 1.5)<0.0001# of ET (mean ± SD)291 (1.9 ± 0.3)494 (2.0 ± 0.4)NS# of embryo vitrified1435 (9.7 ± 7.4)290 (1.1 ± 1.3)<0.0001# of (+)hCG (%)113 (76.3)175 (70.8)NS# of (+)FCA89 (60.1)147 (59.5)NS# of Implantation (%)146 (50.1)219 (44.3)NS Open table in a new tab CONCLUSION: This study demonstrated that high pregnancy and implantation rates were obtained in both fresh and vitrified oocyte donation cycles. With the same clinical outcome as fresh donation cycles, it is possible to use oocyte cryo-banking to minimize the number of oocytes fertilized, thus reduce the number of excess embryos for cryopreservation. As the tremendous demand of oocyte donation remains, the less number of oocyte used per recipient means more recipients could acquire donated oocytes by the use of the oocyte cryo-banking.

OBJECTIVE: Evaluate if the cryopreservation process is associated with a possible increase in the aneuploidy rates in vitrified/warmed oocytes. DESIGN: Prospective, observational study. MATERIALS AND METHODS: The study group included blastocyst stage embryos (donated to research) originated from vitrified oocytes. The control group included blastocysts from a fresh cycle. All cells from each embryo were fixed. Fluorescent in situ hybridization was performed using a 5 chromosome probe mixture (13, 16, 18, 21, and 22). Fisher's exact test was performed, at the level of P<0.05. RESULTS: Ten embryos were obtained from the vitrified oocytes and 16 embryos were obtained from fresh oocytes. A total of 1068 cells were analyzed, 380 in the vitrification group and 688 in the control group. The control group presented a mean of 43.0+/-13.4 cells and the vitrification group 38.0 +/-32.8 cells (NS). The average number of cells with euploid chromosome content was 23.0+/-14.7 (53.5%) in the control group and 20.0+/-28.5 (52.6%) in the vitrification group (NS). In the control group, all embryos were found to be mosaic diploid/aneuploid. Thirteen out of 16 embryos had chaotic distribution. Out of the remaining embryos, 2 had some polyploidy cells; 1 had chromosome 21 monosomy and 1 had chromosome 22 nullisomy/trisomy. The vitrification group, also, all of the embryos were mosaic diploid/aneuploid, however 7 out of the 10 embryos had a chaotic complement and the remaining 3 embryos had haploid content. CONCLUSIONS: Blastocysts obtained from fresh oocytes presented similar cell number as embryos obtained from vitrified oocytes. Also, there was no difference in the distribution of the type and frequency of chromosomal abnormalities, regardless whether it was obtained from fresh or from vitrified oocytes. The results of the current study do not indicate an increase of chromosome abnormality in embryos obtained after oocyte vitrification, further studies including a larger number of embryos is warranted to confirm present findings.

OBJECTIVE: Cryopreserved embryos after slow-freezing are shipped routinely by dry shippers among ART laboratories without affecting their survival rates. However, little information is known about whether the vitrified oocytes/embryos can be maintained in the condition of dry shipper. We aimed to investigate if exposing vitrified oocytes to conditions in a dry shipper used for transportation would impact survival rates.DESIGN: Prospective study.MATERIALS AND METHODS: A total of 163 in-vivo matured MII oocytes from seven donors were cryopreserved on the day of oocyte retrieval and stored in liquid nitrogen dewars as a control group (A). Cryopreservation of oocytes was performed by vitrification using 15% ethylene glycol, 15% DMSO and 0.5 M sucrose with cryotop. The other 70 immature oocytes from the same donors were cultured up to 28 h, and 49 of them were in vitro matured (IVM) and vitrified on the following day of oocyte retrieval. The vitrified IVM oocytes were transferred and held in the dry shipper (MVE sc4/2v) for 60 h as a treatment group (B). Oocyte warming (in both control and treatment groups) was performed by serial dilutions in three steps using 1.0M, 0.5M, and 0M sucrose solutions. Results were analyzed by the Chi-square (P<0.05) test.RESULTS: In group A, a total of 54 oocytes were warmed, and 47 out of 54 survived (87.0%). In group B, 42 out of 49 oocytes survived after vitrification, exposure to vapor phase in dry shipper, and warming (85.7%; NS).CONCLUSIONS: In the present study, we have shown that survival rate of vitrified oocytes was not affected after exposure to the environment of the dry shipper. These results suggest that oocytes cryopreserved by the vitrification technique may be shipped safely using the dry shipper. As an increasing number of IVF clinics are switching from slow-freezing to the vitrification method to cryopreserve oocytes and embryos, the results of this study provide critical information that ensures vitrified oocytes/embryos can be transported in the dry shipper safely. OBJECTIVE: Cryopreserved embryos after slow-freezing are shipped routinely by dry shippers among ART laboratories without affecting their survival rates. However, little information is known about whether the vitrified oocytes/embryos can be maintained in the condition of dry shipper. We aimed to investigate if exposing vitrified oocytes to conditions in a dry shipper used for transportation would impact survival rates. DESIGN: Prospective study. MATERIALS AND METHODS: A total of 163 in-vivo matured MII oocytes from seven donors were cryopreserved on the day of oocyte retrieval and stored in liquid nitrogen dewars as a control group (A). Cryopreservation of oocytes was performed by vitrification using 15% ethylene glycol, 15% DMSO and 0.5 M sucrose with cryotop. The other 70 immature oocytes from the same donors were cultured up to 28 h, and 49 of them were in vitro matured (IVM) and vitrified on the following day of oocyte retrieval. The vitrified IVM oocytes were transferred and held in the dry shipper (MVE sc4/2v) for 60 h as a treatment group (B). Oocyte warming (in both control and treatment groups) was performed by serial dilutions in three steps using 1.0M, 0.5M, and 0M sucrose solutions. Results were analyzed by the Chi-square (P<0.05) test. RESULTS: In group A, a total of 54 oocytes were warmed, and 47 out of 54 survived (87.0%). In group B, 42 out of 49 oocytes survived after vitrification, exposure to vapor phase in dry shipper, and warming (85.7%; NS). CONCLUSIONS: In the present study, we have shown that survival rate of vitrified oocytes was not affected after exposure to the environment of the dry shipper. These results suggest that oocytes cryopreserved by the vitrification technique may be shipped safely using the dry shipper. As an increasing number of IVF clinics are switching from slow-freezing to the vitrification method to cryopreserve oocytes and embryos, the results of this study provide critical information that ensures vitrified oocytes/embryos can be transported in the dry shipper safely.

OBJECTIVE: The ability to efficiently cryopreserve oocytes can serve for various fertility treatments related applications. The aim of this study is to evaluate the applications and results of oocyte cryopreservation in our ART program. DESIGN: Retrospective study. MATERIALS AND METHODS: From May 2006 to April 2009, we have identified 5 distinct egg freezing applications: Group 1 "Rescue" egg cryo cycles when sperm has not been available at egg retrieval, Group 2 Fertility Preservation for single women, Group 3 an elective alternative to embryo freezing when there are ethical concerns with embryo cryopreservation, Group 4 Egg donation program to enhance management of donors and recipients and Group 5 Cancer patients before radio and or chemotherapy. RESULTS: In group 5 there were 14 women (age average 24 ± 4 years old) who have had frozen 220 oocytes (average per patient 14 ± 10) None of those oocytes have been warmed yet. Results are summarized in Table 1.Table 1Efficiency of different applications of oocyte freezing.Groups1234# of Patients82738107Age Avg (+/-SD)33.9 (5.1)35.5 (4.4)33.7 (4.3)26.1 (2.8)# Eggs Avg (mean)105 (13.1)272 (9.7)469 (12.3)2090 (22)# of Patients with Thaw325111Recipient Age avg (+/-SD)34.0 (5.6)35.5 (0.5)34.6 (3.5)41.0 (4.9)# Eggs thawed (mean)37 (12.3)19 (9.5)55 (11)781 (7.0)Survival rate %78846986Fertilization rate %76758289Blastocyst rate %32507469Clinical pregnancy rate %67508065Implantation rate %26173850 Open table in a new tab CONCLUSIONS: The excellent survival, fertilization, blastocyst and pregnancy/implantation rates obtained after egg warming in the different groups demonstrates the high efficiency of oocyte cryo banking when used in combination with a competent vitrification method. These results demonstrate a clear clinical benefit for patients with different medical needs/indications that now can be managed through egg cryopreservation as an advanced treatment option for assisted reproduction.

OBJECTIVE: To evaluate the clinical efficiency of vitrification, at various embryonic development stages, using the Cryolock® device, and to compare it with the slow freezing technique. DESIGN: Retrospective analysis. MATERIALS AND METHODS: A total of 289 thawing cycles of slow freezing and 108 warming cycles of vitrification were performed parallel and data were analyzed. Zygotes and cleaved embryos were frozen and thawed by slow freezing protocol using Freeze-Kit1 and Thaw Kit1 (Vitrolife, Kungsbacka-Sweden) and day-5 embryos using the Blastocyst Freeze / Thaw Kit (SAGE, Biopharma, Trumbull, CT, USA). In the vitrification group, embryos at all stages were vitrified using the same vitrification protocol 15% ethylene glycol, 15% DMSO, and 0.5M sucrose with the Cryolock® (Biodiseno Ltda, Colombia) and warming using series of solutions (1.0M/0.5M sucrose). Results were analyzed using One-way ANOVA and the Fisher's exact tests using P at 0.05 level. Table 1Survival, Pregnancy and Implantation rates after warming/thawing transferred cyclesRatesVitrification %Slow Freezing %P ValueSurvival Rate2PN9785<0.0001Cleaved90750.0044Blastocyst9376<0.0001All9480<0.0001Pregnancy Rate (FCA)2PN5943NSCleaved5043NSBlastocyst6555NSAll61450.0025Implantation Rate/Embryo transf2PN2823NSCleaved3222NSBlastocyst3633NSAll33240.0032Implantation Rate/ Embryo Thawed2PN1410NSCleaved26120.0021Blastocyst3326NSAll2312<0.0001 Open table in a new tab As an additional parameter for embryos at the cleavage cell stage, the number of intact cells (compared to the lysed cells) was also noted. After vitrification, 93% of the original (before freezing) cells remained intact, and after slow freezing 68% (P<0.0001). CONCLUSIONS: The results of this study demonstrate that vitrification at all different stages of human embryo development provides high survival, high pregnancy, implantation rates and many of these outcomes are superior compared to slow-freezing protocol. This study also shows that Cryolock® can function successfully as a carrier device for vitrification in the daily routine laboratory use. These outcomes provide strong evidence on adopting vitrification as a standard protocol for embryo cryopreservation for all developmental stages.

The purpose of this study is to determine if superovulation for IVF with a combination of FSH and LH produces a different outcome when compared to FSH only stimulation. A retrospective data analysis. Patients' cycles were classified into two groups. Group 1 included patients that were given FSH only for stimulation. Group 2 included patients stimulated with FSH in tandem with LH. A total of 1124 consecutive cycles from 996 IVF patients which were attending a private clinic from January 2004 to December 2005. Only donor and recipients were excluded from this study. Patients receiving only FSH were given either Follistim® or Gonal-F ® while patients receiving combination FSH/LH had either Menopur® or Repronex ® with recombinant FSH. Unpaired t-test and chi-square tests were used where appropriate. Nine hundred and ninety seven patient cycles were included in group 1. One-hundred and twenty-seven patient cycles were included in group 2. Patient age was not significantly different between group 1 and group 2, 35.1 ± 4.6 years, 35.6 ± 4.4 years respectively (P=0.2466). Basal FSH was not significantly different between group 1 and group 2, 6.3 ± 2.9 and 6.3 ± 3.8, respectively (P=0.9999). The total number of oocytes retrieved between group 1 (15.4 ± 9.7) and group 2 (12.9 ± 8.8) was significantly different (P=0.0058). Embryo quality on day 3 between group 1 (2715 "A" quality, 2244 "B" quality, 1157 "C" quality, 987 "D" quality) and group 2 (293 "A" quality, 242 "B" quality, 123 "C" quality, 128 "D" quality) was not significantly different (P=0.3409). Pregnancy rates (+hCG) between group 1, 479 of 997 (48.0%), and group 2, 53 of 127 (41.7%) was not significantly different (P=0.1876). Fetal cardiac activity between group 1, 391 of 997 (39.2%), and group 2, 41 of 127 (32.3%), was not significantly different (P=0.1464). Average number of embryos transferred between group 1 (2.3 ± 1.2) and group 2 (2.5 ± 1.3) was not significantly different (P=0.0801). However, implantation rates between group 1, 556/2356 (23.6%), and group 2, 56/321 (17.4%) was significantly different (P=0.0167). FSH in combination with LH yields no advantage in terms of number of oocytes retrieved, embryo quality, or pregnancy rates. This data seems to suggest that implantation rates are lower in group 2 then group 1. This finding may be attributable to the fact that our practice generally resorts to FSH/LH combination protocols in patients with prior IVF failure.

The purpose of this study is to determine if the mode of insemination (ICSI or IVF) has an effect on implantation, using sibling oocytes. Retrospective data evaluation data. Patients were distributed into three groups. Group 1 included patients with only IVF embryos transferred, group 2 included patients with only ICSI embryos transferred (ET), group 3 included patients who had one ICSI and one IVF embryo transferred. Trial 1 consisted of all patients undergoing IVF at a private clinic from January 2004 to December 2005. Inclusion criteria included 1) patients who had half their oocytes inseminated with ICSI and half their oocytes inseminated with IVF, 2) only had 2 embryos transferred on day 3 and, 3) did not experience failed fertilization for either ICSI or IVF. Trial 2 included the same patients as in trial 1, however, patients which only had ICSI or IVF transferred due poor embryo quality in the other group were excluded. Therefore, in this trial, the embryologist had the opportunity to select either an ICSI and/or IVF embryo for transfer. The unpaired t-test and chi-square tests were used where appropriate. For trial 1, the average age and embryo quality at ET between the three groups was not significant. Pregnancy rates (+hCG) between group 1, 2, and 3, was not significantly different, 57 of 75 (76.0%), 40 of 55 (72.7%), and 68 of 96 (70.8%), respectively (P=0.7508). Fetal cardiac activity (+FCA) between group 1, 2, and 3 was not significantly different, 48 of 75 (64.0%), 36 of 55 (65.4%), 60 of 96 (62.5%), respectively (P=0.9343). Implantation rates between group 1 (67/150), group 2 (47/110), and group 3 (80/192) was not significantly different (P=0.8557). For trial 2, the average age and embryo quality at ET between the three groups was not significantly different. Pregnancy rates (+hCG) between group 1, 2, and 3, was not significantly different, 37/44 (84.1%), 27/37 (72.9%), and 68/96 (70.8%) respectively (P=0.2393). Fetal cardiac activity (+FCA) between group 1, 2, and 3 was not significantly different, 32 of 44 (72.7%), 25 of 37 (67.6%), and 60 of 96 (62.5%), respectively (P=0.4835). Implantation rates between group 1 (45/88), group 2 (34/70), and group 3 (80/192) was also not significantly different (P=0.2820). The results of our study suggest that implantation rates of sibling embryos derived from either IVF or ICSI insemination does not differ. The type of insemination should not play a role when choosing embryos for transfer.

The use of aneuploidy screening (PGD-AS) is rapidly growing. To obtain oocytes, various ovarian stimulation protocols are used; however, information is lacking if any of those protocols have particular advantage in regard to embryonic chromosomal normality, as it may be reflected by aneuploidy screening. To answer this question, we aimed to analyze aneuploidy rates in embryos obtained from IVF patients submitted to different ovarian stimulation protocols. To minimize the effect of age, we have selected patients with age of less than 35 years. Retrospective. Ovarian stimulation was performed using: 1, Antagonist protocol with rFSH alone (total of 50 patients; Group-1); 2, Lupron down-regulation with rFSH alone (40 patients; Group-2) or 3, Flare protocol with rFSH alone (27 patients; Group-3; study period: 2004–2006). Indications of PGD: Implantation failure, pregnancy loss, unexplained infertility – incidences were similar in all three groups. Embryos were biopsied on day 3; fixed cells were tested with 9 chromosomal probes. Data on embryo development/grade, aneuploidy rates, and implantation rates were compared between the three groups. One-way ANOVA and Chi-square tests were used for statistical analyses; P<0.05 was considered to be significant. Average age of women were 32.1; 32.4 and 32.8 in Groups 1 to 3 respectively (NS). A total of 985, 895 and 359 oocytes were obtained in Groups 1, 2, 3 respectively (P<0.01 between Group 1 vs. Group 2 and 3). Fertilization rates were 57%, 58% and 58% in the three groups (NS). Incidence of excellent plus good quality embryos were significantly higher in Group 1 and 2 compared to Group 3 (75%, 73% and 67% respectively, P<0.05). Rate of chromosomally abnormal embryos (aneuploidy rates) were: 59%, 64% and 63% in Groups 1 to 3 respectively (NS). Embryo implantation rates were 24%, 27% and 30% in the three groups respectively (NS). The results demonstrate that frequency of aneuploid embryos is similarly high in young infertile patients, regardless to the type of ovarian stimulation protocol used. Compromised embryo development was more frequently observed in Group-3 (Lupron short/flare protocol), however, this was not reflected in PGD or implantation outcomes. As a conclusion, we may say that all three ovarian stimulation protocols provided comparable laboratory and clinical outcomes and none were associated with particular advantage or disadvantage in regard to aneuploidy or implantation results.