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Blastocyst culture selects for euploid embryos: comparison of blastomere and trophectoderm biopsies

Adler, Alexis; Lee, Hsaio-Ling; McCulloh, David H; Ampeloquio, Esmeralda; Clarke-Williams, Melicia; Wertz, Brooke Hodes; Grifo, James
Preimplantation genetic diagnosis and screening improves the chances of achieving a viable pregnancy, not only free of undesired single-gene defects but also aneuploidy. In addition, improvements in vitrification provide an efficient means of preserving embryos (blastocysts). By combining trophectoderm biopsy with recent improvements in vitrification methods, only those embryos that have proved themselves viable and potentially more competent are tested. Using array comparative genomic hybridization (aCGH) to assess all 24 chromosomes, aneuploidy rates were compared between day-3 blastomere biopsy and day-5 trophectoderm biopsy. Of those 1603 embryos, 31% were euploid, 62% were aneuploid and 7% not analysable. A significantly larger proportion of embryos were euploid on day-5 biopsy (42%) compared with day-3 biopsy (24%, P<0.0001). The number of euploid embryos per patient was not significantly different. Combining extended culture, trophectoderm biopsy and aneuploidy assessment by aCGH and subsequent vitrification can provide a more efficient means of achieving euploid pregnancies in IVF. Preimplantation genetic diagnosis and screening improves the chances of achieving a viable pregnancy, not only free of undesired single-gene defects but also aneuploidy. In addition, improvements in vitrification provide an efficient means of preserving embryos (blastocysts). By combining trophectoderm biopsy with recent improvements in vitrification methods, only those embryos that have proved themselves viable and potentially more competent are tested. Using array comparative genomic hybridization to assess all 24 chromosomes, aneuploidy rates were compared between day-3 blastomere biopsy and trophectoderm biopsy. Of those 1603 embryos, 31% were euploid, 62% were aneuploid and 7% not analysable. A significantly larger proportion of embryos were euploid with trophectoderm biopsy (42%) compared with blastomere biopsy (24%, P<0.0001). The number of euploid embryos per patient was not significantly different. Combining extended culture, trophectoderm biopsy and aneuploidy assessment by array comparative genomic hybridization and subsequent vitrification can provide a more efficient means of achieving euploid pregnancies in IVF.
PMID: 24581980
ISSN: 1472-6483
CID: 929922

Diminished Effect of Maternal Age on Implantation After Preimplantation Genetic Diagnosis With Array Comparative Genomic Hybridization [Editorial]

Harton, Gary L; Munne, Santiago; Surrey, Mark; Grifo, Jamie; Kaplan, Brian; McCulloh, David H; Griffin, Darren K; Wells, Dagan; PGD Practitioners Grp
The chief cause for failure of in vitro fertilization (IVF)-assisted reproductive treatments may be the high frequency of aneuploid preimplantation embryos, especially among women of advanced reproductive age. It has been hypothesized that pregnancy loss with advancing maternal age can be prevented by selective transfer of euploid embryos. Preimplantation genetic diagnosis (PGD) for aneuploidy was first attempted more than 20 years ago. It was hoped that screening embryos for aneuploidy and transferring only those found to be euploid would increase implantation and pregnancy rates and reduce pregnancy loss rates. Initial attempts to detect aneuploidy used fluorescence in situ hybridization analysis (first-generation PGD). However, several randomized controlled trials found no benefit for first-generation PGD or even a negative impact on implantation, pregnancy, or loss rates. A more accurate version of PGD was needed. Array comparative genomic hybridization (aCGH) is a second-generation PGD. Use of this technique in randomized controlled trials improved pregnancy rates. The major drawback of aCGH is the need for at least 3 full days for an analysis to be completed. With the introduction of vitrification (freezing), safe cryopreservation of embryos that underwent biopsy became possible, allowing delay of embryo biopsy from 3 days (gastrula stage) to 5 to 6 days (blastocyst stage), which is less detrimental to embryo development. This multicenter retrospective study assessed the relationship between maternal age, chromosome abnormality, implantation, and pregnancy loss. The aim of the study was to determine whether aCGH followed by selective transfer of euploid embryos would mitigate the age-related decline in implantation rates observed in IVF cycles. Preimplantation chromosome screening was performed in women undergoing IVF at a number of fertility clinics in the United States. Implantation rates across different maternal ages were examined with embryo biopsy on day 3 or day 5/6 followed by aCGH. The primary outcome measures were aneuploidy, implantation, pregnancy, and loss rates. Aneuploidy rates increased with advancing maternal age from 53% to 93% for day 3 biopsies and from 32% to 85% for day 5/6 biopsies. Implantation rates for euploid embryos for ages 35 to 42 years were maintained after PGD; rates ranged from 44% to 32% for day 3 biopsies and 51% to 40% for day 5 biopsies. Ongoing pregnancy rates per transfer remained stable for maternal ages younger than 42 years, ranging from 48.5% to 38.1% for day 3 biopsies and 64.4% to 54.5% for day 5 biopsies. For patients 42 years or older, implantation rates were 23.3% with day 3 biopsies and 27.7% with day 5/6, and the ongoing pregnancy rate was 9.3% for day 3 biopsies and 10.3% for day 5. These data show no significant difference in implantation and pregnancy rates after selective transfer of euploid embryos between reproductively younger and older patients up to 42 years old. The enhanced embryo selection afforded by methods such as aCGH cannot improve pregnancy rates in patients who fail to produce at least 1 euploid embryo, a situation increasingly common with advancing maternal age. These findings and mounting data from other studies suggest that aneuploidy is the primary reason for the marked decline in IVF treatment success rates in women of advanced reproductive age when PGD is not used.
ISI:000346280900017
ISSN: 1533-9866
CID: 2338562

Diminished effect of maternal age on implantation after preimplantation genetic diagnosis with array comparative genomic hybridization

Harton, Gary L; Munne, Santiago; Surrey, Mark; Grifo, Jamie; Kaplan, Brian; McCulloh, David H; Griffin, Darren K; Wells, Dagan
OBJECTIVE: To assess the relationship between maternal age, chromosome abnormality, implantation, and pregnancy loss. DESIGN: Multicenter retrospective study. SETTING: IVF centers in the United States. PATIENT(S): IVF patients undergoing chromosome screening. INTERVENTION(S): Embryo biopsy on day 3 or day 5/6 with preimplantation genetic diagnosis (PGD) by array comparative genomic hybridization. MAIN OUTCOME MEASURE(S): Aneuploidy, implantation, pregnancy, and loss rates. RESULT(S): Aneuploidy rates increased with maternal age from 53% to 93% for day 3 biopsies and from 32% to 85% for blastocyst biopsies. Implantation rates for euploid embryos for ages <35-42 years did not decrease after PGD: ranges 44%-32% for day 3 and 51%-40% for blastocyst. Ongoing pregnancy rates per transfer did not decrease for maternal ages <42 years after PGD with day 3 biopsy (48.5%-38.1%) or blastocyst biopsy (64.4%-54.5%). Patients >42 years old had implantation rates of 23.3% (day 3), 27.7% (day 5/6), and the pregnancy rate with day 3 biopsy was 9.3% and with day 5 biopsy 10.3%. CONCLUSION(S): Selective transfer of euploid embryos showed that implantation and pregnancy rates were not significantly different between reproductively younger and older patients up to age 42 years. Some patients who start an IVF cycle planning to have chromosome screening do not have euploid embryos available for transfer, a situation that increases with advancing maternal age. Mounting data suggests that the dramatic decline in IVF treatment success rates with female age is primarily caused by aneuploidy.
PMID: 24034939
ISSN: 0015-0282
CID: 929912

Live birth from previously vitrified oocytes, after trophectoderm biopsy, revitrification, and transfer of a euploid blastocyst

Grifo, Jamie A; Hodes-Wertz, Brooke; Lee, Hsiao Ling; Ampeloquio, Esmeralda; Clarke-Williams, Melicia; Adler, Alexis; Munne, Santiago; Berkeley, Alan S
Our objective is to describe a successful live birth from oocyte vitrification followed by thaw, fertilization, blastocyst culture, trophectoderm biopsy, vitrification, and subsequent thaw. Fifteen mature oocytes were frozen from a patient with uterine factor infertility. Thirteen oocytes survived the thaw, and five underwent trophectoderm biopsy and were refrozen. Three euploid embryos were obtained. A single euploid embryo was transferred in the second thaw cycle to a known recipient leading to the delivery of a normal male infant. This case report is proof of the concept that preimplantation screening and diagnosis is an option for fertility preservation patients.
PMCID:3888074
PMID: 24453522
ISSN: 1179-5581
CID: 760042

Oocyte efficiency: does live birth rate differ when analyzing cryopreserved and fresh oocytes on a per-oocyte basis?

Goldman, Kara N; Noyes, Nicole L; Knopman, Jaime M; McCaffrey, Caroline; Grifo, James A
OBJECTIVE: To compare the efficiency of oocyte cryopreservation (OC) and IVF using the metric "live births per mature oocyte retrieved." DESIGN: Retrospective analysis. SETTING: University-based fertility center. PATIENT(S): Forty women who underwent OC with thaw attempt between 2004 and 2010; 25 autologous and 15 donor-oocyte treatments were included. One thousand nine hundred eight women underwent their first, fresh conventional IVF treatment between 2004 and 2010; 1,392 used autologous oocytes, and 516 used donor oocytes. Autologous and donor-oocyte cycles were analyzed separately. All oocytes were obtained from women
PMID: 23721713
ISSN: 0015-0282
CID: 519402

Clinical outcomes of natural versus medicated frozen-thawed embryo transfers (FET): A 7-year review [Meeting Abstract]

Nazem, T G; Hodes-Wertz, B; Davison, J Z; Grifo, J A
OBJECTIVE: To compare cycle and clinical outcomes between natural or medicated FET cycles. DESIGN: Retrospective study in an academic institution. MATERIALS AND METHODS: There were 1177 FET cycles from 2005- 2012. FET included embryos created from autologous or donor oocytes. Embryo transfers from oocyte thaws were excluded. Groups were analyzed for % donor cycles, % vitrification, % preimplantation genetic diagnosis/ screening (PGD), number of monitoring days, embryos transferred (ET), embryos thawed, day of embryo transfer, maximal endometrial (EM) thickness before transfer, peak estradiol (E2) and progesterone (P4) in follicular cycle, clinical pregnancy rate (CPR), spontaneous abortion rate (SABR) and ongoing/live birth rate (OP/LBR). CPR was defined by fetal cardiac activity on 1st-trimester ultrasound. SABR was calculated per pregnancy with a sac. T-tests were performed with p<0.05. RESULTS: Subset analysis was performed excluding PGD cycles, confirming no difference between natural and medicated cycles except for days of monitoring, peak E2 and peak P4. CONCLUSION: There can be advantages to both natural FET (lack of medication costs and injections) and medicated cycles (flexibility). Natural and medicated FET are equally effective in terms of pregnancy outcomes and should be offered based on patient preference and suitability (Table Presented)
EMBASE:71164388
ISSN: 0015-0282
CID: 549982

An analysis of comprehensive carrier screening results for 1000 clinical samples: The importance of transparent disease classification & criteria for selection [Meeting Abstract]

Bisignano, A; Kumar, N; Prates, R N; Munne, S; Grifo, J; Hoffman, D
OBJECTIVE: Advancements in genomics allow for expanded, cost-effective, and high-throughput carrier screening. The American College of Medical Genetics (ACMG) recommends that phenotype and penetrance should be considered in selecting diseases for inclusion. Based on 1000 clinical samples screened, the selected diseases were classified according to these factors with the goal of improving transparency, pre- and post-test counseling and results disclosure protocols. DESIGN: Retrospective study. MATERIALS AND METHODS: Recombine's Comprehensive Carrier Screen for 978 mutations associated with 181 recessive diseases was performed on 1000 clinical referrals. Diseases were classified into 2 groups: high impact (significant effect on quality of life/reduced lifespan) and variable spectrum (less severe phenotype/lower penetrance). Informed consent to utilize de-identified data was obtained from all patients. RESULTS: See table (Table Presented) CONCLUSION:With the full panel, ~99% of patients carry at least 1 mutation. More than 50% of samples carry MTHFR mutations. Considering only high-impact diseases, ~40% of patients carry at least 1 mutation. Inclusion of mild disorders may create anxiety and logistical concerns for patients and physicians. This anxiety can be reduced with post-test genetic counseling, transparency about differences in severity between high-impact and variable spectrum diseases in all materials (pre-counseling, web-based & reports), and the ability to customize the disease panel
EMBASE:71163959
ISSN: 0015-0282
CID: 549992

Is cohort size of euploid blastocysts following comprehensive chromosomal screening predictive of improved outcomes in single embryo transfer cycles [Meeting Abstract]

Morin, S.; Melzer, K.; Grifo, J.; Colls, P.; Zheng, Z.; Munne, S.
ISI:000320467700113
ISSN: 0268-1161
CID: 427402

A Comparison of Pregnancy Outcomes between Day 3 and Day 5 Embryo Transfers: Is There More to the Story than SART? [Meeting Abstract]

McClelland, Susan; Melzer, Katherine; McCulloh, David; Grifo, James A.
ISI:000315281800008
ISSN: 0015-0282
CID: 249222

Identification of Ideal Candidates for Single Embryo Transfer (SET) of a Euploid Blastocyst Based upon Overall Embryo Cohort Euploidy Rate [Meeting Abstract]

Morin, Scott; Melzer, Katherine; Grifo, Jamie; Colls, Pere; Zheng, Xyezhong; Munne, Santiago
ISI:000329543100580
ISSN: 1933-7191
CID: 808002