ObjectiveThe objective of this study was to determine if embryos derived from vitrified donor oocytes perform in a similar manner to oocytes derived from fresh cycles after a second round of vitrification and warming (ie FET cycles).DesignA retrospective analysis of all donor FET cycles from January 2011 through December 2013. The initial retrievals and oocyte warming took place from June 2009 to December 2013. Group A are cycles in which donor oocytes were retrieved, inseminated, cultured to the blastocyst stage, vitrified then warmed at a later time for an ET. Group B are cycles in which donor oocytes (obtained from the Fairfax EggBank) were first vitrified and later warmed, inseminated, cultured to the blastocyst stage, then vitrified again for future FET. The endpoints utilized for this study are embryo survival, implantation and clinical pregnancy outcome.Materials and MethodsOvarian stimulation and oocyte retrievals were performed using standard protocols. Oocyte vitrification was performed on cryotops (Kitazato, Japan), blastocyst vitrification was performed using Cryotips (Irvine Scientific, USA). Both oocytes and embryos were vitrified using ethylene glycol and DMSO as cryoprotectants in gradually increasing amounts. Blastocysts were collapsed prior to vitrification using a single laser pulse (Xylos). Warming protocols were accomplished using rapid warming first at 37 C, then a gradual return to isotonic media at room temperature.ResultsTable 1Comparison of Group A and Group B# Cycles# Embs VitrifiedEmbs SurvivedEmbs/Transfer# ImplantationClin PregnancySABGroup A236404388 (96%)1.6137 (35%)119 (51%)11 (9%)Group B *213837 (97%)1.816 (43%)12 (57%)1 (8%)* Frozen Oocytes from Fairfax Egg Bank Open table in a new tab ConclusionVitrifying and warming oocytes has now become common and is gaining in popularity as an alternative and effective method of delivering donor egg services to patients. Results across the globe are similar to results in fresh donor egg cycles. This has led to the establishment of commercial egg banks. One of the remaining questions is whether or not these previously frozen oocytes, once warmed, inseminated and cultured to later embryonic stages can be again re-vitrified and warmed with similar results as in fresh donor egg cycles. This data set suggests that the laboratory and clinical results are comparable and quite good in both groups. ObjectiveThe objective of this study was to determine if embryos derived from vitrified donor oocytes perform in a similar manner to oocytes derived from fresh cycles after a second round of vitrification and warming (ie FET cycles). The objective of this study was to determine if embryos derived from vitrified donor oocytes perform in a similar manner to oocytes derived from fresh cycles after a second round of vitrification and warming (ie FET cycles). DesignA retrospective analysis of all donor FET cycles from January 2011 through December 2013. The initial retrievals and oocyte warming took place from June 2009 to December 2013. Group A are cycles in which donor oocytes were retrieved, inseminated, cultured to the blastocyst stage, vitrified then warmed at a later time for an ET. Group B are cycles in which donor oocytes (obtained from the Fairfax EggBank) were first vitrified and later warmed, inseminated, cultured to the blastocyst stage, then vitrified again for future FET. The endpoints utilized for this study are embryo survival, implantation and clinical pregnancy outcome. A retrospective analysis of all donor FET cycles from January 2011 through December 2013. The initial retrievals and oocyte warming took place from June 2009 to December 2013. Group A are cycles in which donor oocytes were retrieved, inseminated, cultured to the blastocyst stage, vitrified then warmed at a later time for an ET. Group B are cycles in which donor oocytes (obtained from the Fairfax EggBank) were first vitrified and later warmed, inseminated, cultured to the blastocyst stage, then vitrified again for future FET. The endpoints utilized for this study are embryo survival, implantation and clinical pregnancy outcome. Materials and MethodsOvarian stimulation and oocyte retrievals were performed using standard protocols. Oocyte vitrification was performed on cryotops (Kitazato, Japan), blastocyst vitrification was performed using Cryotips (Irvine Scientific, USA). Both oocytes and embryos were vitrified using ethylene glycol and DMSO as cryoprotectants in gradually increasing amounts. Blastocysts were collapsed prior to vitrification using a single laser pulse (Xylos). Warming protocols were accomplished using rapid warming first at 37 C, then a gradual return to isotonic media at room temperature. Ovarian stimulation and oocyte retrievals were performed using standard protocols. Oocyte vitrification was performed on cryotops (Kitazato, Japan), blastocyst vitrification was performed using Cryotips (Irvine Scientific, USA). Both oocytes and embryos were vitrified using ethylene glycol and DMSO as cryoprotectants in gradually increasing amounts. Blastocysts were collapsed prior to vitrification using a single laser pulse (Xylos). Warming protocols were accomplished using rapid warming first at 37 C, then a gradual return to isotonic media at room temperature. ResultsTable 1Comparison of Group A and Group B# Cycles# Embs VitrifiedEmbs SurvivedEmbs/Transfer# ImplantationClin PregnancySABGroup A236404388 (96%)1.6137 (35%)119 (51%)11 (9%)Group B *213837 (97%)1.816 (43%)12 (57%)1 (8%)* Frozen Oocytes from Fairfax Egg Bank Open table in a new tab * Frozen Oocytes from Fairfax Egg Bank ConclusionVitrifying and warming oocytes has now become common and is gaining in popularity as an alternative and effective method of delivering donor egg services to patients. Results across the globe are similar to results in fresh donor egg cycles. This has led to the establishment of commercial egg banks. One of the remaining questions is whether or not these previously frozen oocytes, once warmed, inseminated and cultured to later embryonic stages can be again re-vitrified and warmed with similar results as in fresh donor egg cycles. This data set suggests that the laboratory and clinical results are comparable and quite good in both groups. Vitrifying and warming oocytes has now become common and is gaining in popularity as an alternative and effective method of delivering donor egg services to patients. Results across the globe are similar to results in fresh donor egg cycles. This has led to the establishment of commercial egg banks. One of the remaining questions is whether or not these previously frozen oocytes, once warmed, inseminated and cultured to later embryonic stages can be again re-vitrified and warmed with similar results as in fresh donor egg cycles. This data set suggests that the laboratory and clinical results are comparable and quite good in both groups.

To study the developmental impact of embryo biopsy on the progression of embryos from day 3 to day 5. Retrospective analysis of IVF cycles from January 2006 through December 2006. During this time period, 105 patients (Group 1) had embryo biopsy for preimplantation genetic diagnosis (PGD) and 253 patient cycles were cultured to day 5 for blastocyst transfer (Group 2) without biopsy. Indications for PGD were: aneuploidy screening, chromosome rearrangements, family balancing, and various single gene defects. Ovarian stimulation and IVF were done with standard methods. Quinn's sequential media (Sage) was used throughout the study. Embryos were cultured in individual 20 μl droplets under oil in 5% CO2 and 5% O2. Biopsy was done either by physical slit or laser heat ablation of the zona pellucida. A single cell was removed from each embryo that had progressed to at least 5 cells on the morning of day 3 of embryo development. Embryos were grouped and compared by their cleavage stage of development on Day 3 and their progression to blastocyst. An embryo was considered a blastocyst if it formed a blastoceol and an inner cell mass (ICM) could be clearly delineated. Chi-squared analysis was done comparing the outcome of the various groups and p values are displayed in Table 1. A p value of < 0.05 was considered statistically significant. 781 embryos were cultured and followed in Group 1 (Biopsy) and 1,959 embryos were cultured and followed in Group 2 (No Biopsy). The results of the study can be seen in Table 1 below: TableDay 5 Group 1 (Biopsy)Day 5 Group 2 (No Biopsy)Stage D3NBlasts% BlastsNBlasts% BlastsP5 Cells117762656725<0.0016 Cells139312236713236<0.057 Cells191743943020648N.S.8 Cells2421174868641360N.S.9+ Cells77476121113563N.S.Total78127335195995349<0.001 Open table in a new tab Performing single blastomere embryo biopsy on Day 3 embryos clearly had a negative impact on progression to the blastocyst stage on Day 5. However, the effects were more pronounced on embryos that were already progressing slowly. For embryos at later cell stages (at least 7 cells or higher) the impact was minimal and did not reach statistical significance. This information may be used counseling patients considering embryo biopsy.

In some situations where embryo biopsy and Pre-Implantation Genetic Diagnosis (PGD) are being performed, it is necessary to freeze some or all of the embryos created in the fresh IVF cycle. These clinical situations are: 1) Thin endometrial lining, 2) Ovarian Hyper Stimulation Syndrome (OHSS), or 3) Excessive embryos are created. Our purpose is to determine if cryopreserving embryos prior to biopsy and PGD is a feasible clinical alternative, and whether developmental stage has an impact on the outcome. A retrospective study comparing the survival rates of embryos and the pregnancy rates of patients in which embryos were cryopreserved at either the pronucleate stage or at early cleavage stages and then later biopsied for PGD. All embryos were cryopreserved using standard controlled rate freezing and slow thawing methods. Embryos were individually cryopreserved and stored in straws. Mechanical biopsy was performed on day 3 on all embryos with ≥5 blastomeres. When possible, 2 blastomeres were removed for genetic diagnosis. A total of 171 pronuclear zygotes and 214 cleaved embryos were utilized from 23 and 29 patients respectively. Table 1 shows the number of thaw cycles, numbers of biopsies and transfers and the outcomes of the thaws at each developmental stage. There was no significant difference in any of these parameters among the 3 groups. Table 1Tx = Transfer Bx = Biopsy Clinical Pregnancy = Fetal Heart Beat on Ultrasound Tx = Transfer Bx = Biopsy Clinical Pregnancy = Fetal Heart Beat on Ultrasound Embryos from day 1, day 2 and day 3 were all used successfully. The most effective and efficient method from a logistical standpoint is to freeze embryos on day 1 or day 2, then thaw on the equivalent day in a subsequent frozen embryo transfer cycle prior to doing biopsy and genetic diagnosis. We conclude that embryos can be successfully frozen, thawed and biopsied when it is clinically necessary or advisable to do so; with very reasonable results.

Purpose: To determine whether donor oocyte cytoplasm transferred into the oocytes of women ≥40 years or with diminished ovarian reserve would enhance embryo quality, implantation, or pregnancy rates.

Objectives: IVF is an unquestionably important treatment for infertile couples. An undesirable but unavoidable risk of cotransferring several embryos is multiple gestations. Avoidance of all dizygotic multiple gestations cannot be achieved without a substantial reduction in cycle success rates. We evaluated our FET data for factors affecting the frequency of high-order multiple gestations. Design: Retrospective study design from a large private infertility center. Methods: Retrospective review of 1380 consecutive frozen-thawed non-donor embryo transfer cycles after conventional IVF or ICSI and 402 after donor IVF cycles. The number of 100%-intact embryos transferred was the most significant variable predicting a live-born delivery (stepwise logistic regression) and this variable was used to evaluate the probability of high-order multiple gestations. Results: 186 pregnancies resulted in delivery of one or more infants after 1380 non-donor FET cycles; and 90 followed 402 donor FET cycles. No high-order multiple gestation occurred with ≤2 100%-intact embryos in non-donor cycles or with <2 100%-intact embryos in donor cycles regardless of the number of additional <100%-intact embryos transferred. No high-order multiple births occurred when only 2 donor 100%-intact embryos were transferred alone. Donor cycles were more likely to result in pregnancy and high-order multiple births (p=0.04). legend% Embryos transferred with 100% blastomere survivallegend∗ In many cases one or more embryos with <100%-intact blastomeres were also transferred., legend∗∗ Only one quadruplet pregnancy occurred.Non-donorDonorNumber of cyclesLive- born pregTriplets+ (%) of preg)Number of cyclesLive- born preg∗∗Triplets+ (% of preg)02238.1%0%6116.4%0%14058.6%0%10512.4%0%240914.7%0%11823.7%10.7%326719.1%3.9%9328.0%3.7%46729.9%10.0%2343.5%20.0%5922.2%0%250.0%0%Total138013.5%2.1%40221.9%6.7%legend ∗ In many cases one or more embryos with <100%-intact blastomeres were also transferred.legend ∗∗ Only one quadruplet pregnancy occurred. Open table in a new tab Conclusions: Our data indicates that limiting the number of 100%-intact embryos transferred in FET cycles will decrease high-order multiple gestations particularly in donor FET cycles. For those couples who are particularly risk adverse, limiting non-donor FET cycles to ≤2 100%-intact embryos and donor FET cycles to one 100%-intact embryos plus any <100% embryos or 2 100%-intact embryos only will almost completely avoid the risk of high-order multiple gestation but at the expense of a reduction in liveborn infants. Overall, the risk of a high-order multiple gestations is well under 10% for both non-donor and donor FET cycles.