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Assisted Hatching of the Zona Pellucida Prior to Embryo Transfer in Fresh Donor Egg IVF Cycles
RESULT(S): The EDICT contains a section to assess objective understanding and a subjective section to assess donors’ perceived understanding. The objective portion of the EDICT consists of 28 true/false questions derived from the HHS eight basic elements of informed consent. Questions cover the process of oocyte donation, potential risks, legal, and psychological issues. An example question is, ‘‘There is no chance that I can become pregnant during the process of egg donation’’. We found there to be satisfactory internal consistency using Cronbach’s alpha. CONCLUSION(S): At present, there is no standardized method used to counsel oocyte donors, nor is there a mechanism to assess the adequacy of the informed consent process for oocyte donation. For the first time, we describe development of a tool for assessing oocyte donors’ understanding of the donation process and its associated risks. Here we present our ongoing work aiming to validate and test the utility of the EDICT in assessing oocyte donors’ understanding. SUPPORT: None
P-65 Oocyte Defects Leading to Triploid Miscarriages After Donor Oocyte IVF. F.E. Marshall,a B. Behr,b R.B. Lathi.b a Department of Ob/Gyn, University of California Irvine Medical Center, Orange, CA; b Stanford Center for Reproductive Medicine, Stanford, Palo Alto, CA. BACKGROUND: The most common form of aneuploidy observed in first trimester miscarriages is trisomy, followed by polyploidy. Whereas, the risk of meiotic non-disjunction leading to trisomy increases with maternal age, the risk factors for triploid conceptions are poorly understood. Diandric triploidy is commonly thought to be due to polyspermia, but the etiology of and risk factors for digynic triploidy are unknown. OBJECTIVE(S): We present two cases of digynic triploidy after IVF with donor oocytes, leading to subsequent miscarriage. MATERIALS AND METHOD(S): Detailed chart reviewed two cases of triploid pregnancies after IVF cycle with donor oocytes. Both 26 year- old donors underwent controlled ovarian stimulation protocol using long Lupron with 75 IU rFSH and 75 IU hMG for 10 days. After adequate follicular response, 10,000 IU hCG was administered, and oocyte retrieval was performed 35 hours later. Oocytes were inseminated with approximately 25,000 motile sperm/ml and fertilization was evaluated 16-18 hours later. We evaluated IVF cycle parameters and sent miscarriage tissue for SNP microarray analysis. RESULT(S): Case 1: Peak E2 reached 4,375.7 and 30 oocytes retrieved. Post wash semen parameters: 41 M/ml with 91% motility and standard insemination performed. On day 1, three 2PNs were noted, four 3PN, and twenty-three non-fertilized. Embryologist noted poor oocyte quality and faint PNs. On day 3, the best two embryos were transferred: an 8 cell (grade I) from an embryo where no pronuclei were seen and a 6 cell (grade II) from a 2PN. Case 2: Peak E2 reached 3342, and 19 oocytes retrieved. Post wash semen parameters: 37 M/ml with 89% motility. On day 1, four 2PNs noted, and eleven non-fertilized. On day 3, the best two embryos were transferred: two 8 cell (grade II and compacting). The non-transferred embryos did not develop into blastocysts. Ultrasounds confirmed embryonic demise by 8 weeks. Chromosome analysis using SNP microarray revealed triploidy 69 XXX - maternal origin. CONCLUSION(S): These 2 cases illustrate that triploidy of maternal origin can cause miscarriage after oocyte donation. Our hypothesis is that low fertilization rate in the setting of normal semen parameters was a sign of poor oocyte quality. Therefore, we recommended to both couples that they consider a new oocyte donor for subsequent pregnancy attempts. Further studies are needed to evaluate the impact of patient characteristics, stimulation protocol, ovulation induction, and culture conditions on oocyte defects leading to triploidy.
P-66 Assisted Hatching of the Zona Pellucida Prior to Embryo Transfer in Fresh Donor Egg IVF Cycles. L. Cieslik, A. Khabani, C. Khabani, R. Dunsmoor-Su, J. Lamb, L. Hickok. Pacific NW Fertility, Seattle WA; University of Washington, Dept of Obstetrics and Gynecology, School of Medicine, Seattle WA.
S32
Abstracts
BACKGROUND: The early embryo is surrounded by the zona pellucida (ZP). Prior to implantation, the blastocyst must ‘‘hatch’’ out of the ZP so that implantation can occur. A variety of factors may influence the ability of the blastocyst to accomplish hatching and if so influence the chances for pregnancy occurring. Assisted hatching in in vitro fertilization cycles may improve the likelihood of implantation and pregnancy in certain age groups but it has not been well described in a donor/recipient IVF population. OBJECTIVE(S): The purpose of the study was to evaluate the effectiveness of day 5 assisted hatching of the zona pellucida prior to fresh embryo transfer in donor recipients. MATERIALS AND METHOD(S): This study was a retrospective casecontrol analysis of patients who underwent a fresh IVF cycle using donor eggs. The control group of 192 patients had embryos transferred prior to the implementation of assisted hatching of the ZP at our center in 2009 and 127 patients had an embryo transfer after assisted hatching of the ZP with CO2 laser during 2010. Cases and controls were compared using ttest and chi squared analyses. RESULT(S): There were no significant differences in age (41.5 vs 41.4 years), race, estradiol levels of the donor at the time of hCG administration (4130pg/ml vs 3965pg/ml), number of oocytes retrieved (23.5 vs 23.9) or M2s undergoing itracytoplasmic sperm injection (ICSI) (18.3 vs 18.9), number of oocytes fertilized (15.0 vs 16.1), blastocyst formation (9.2 vs 9.5) or the number of embryos transferred (1.66 vs 1.6). There was no difference in the biochemical pregnancy rate (78.7% vs 79.7%) or the number of gestational sacs (1.17 vs1.14). There was a non-statistically significant trend toward an increased probability of observing a fetal heartbeat (78.7% vs 76.6%, P¼0.65) but a trend toward an increase in live birth (75.6% vs 69.8%, P¼0.25) in cases versus controls. (Table1)
TABLE 1.
Chemical pregnancy rate Number of gestational sacs Clinical pregnancy rate Live birth or continuing pregnancy
Hatched (n¼127)
Controls (n¼192)
P value
78.7% 1.17 78.7% 75.6%
79.7% 1.14 76.7% 69.8%
0.83 0.70 0.65 0.25
CONCLUSION(S): This study is the first to evaluate the use of assisted hatching in donor egg IVF cycles. Day 5 Laser-assisted hatching of the zona pellucida prior to embryo transfer in donor IVF cycles may improve clinical pregnancy and live birth rates. FINANCIAL SUPPORT: none
P-67 Does Adenomyosis Impair Implantation? A Cohort Study. Z. Khan, MBBS,a D.M. Breitkopf, MD,b M. Al Hilli, MBBCh,b J.R. Jensen, MD,a C.C. Coddington, MD,a E.A. Stewart, MDa a Division of Reproductive Endocrinology & Infertility, Department of Obstetrics & Gynecology, Mayo Clinic, Rochester, MN 55905; b Department of Obstetrics & Gynecology, Mayo Clinic, Rochester, MN 55905. OBJECTIVE(S): The clinical impact of adenomyosis on infertility is controversial. The aim of this study was to determine the effect of ultrasound-diagnosed adenomyosis on implantation rates with good quality embryos in women undergoing IVF. MATERIALS AND METHOD(S): With IRB approval, all women undergoing fresh and frozen embryo transfers with good quality embryos (embryos at appropriate stage of development with minimal fragmentation) at Mayo Clinic, Rochester from January 2009 to December 2010 were included. Images from pre-IVF ultrasounds performed by the same ultrasonographer were reviewed independently by two physicians blinded to cycle outcome. Adenomyosis was diagnosed only when at least three of the five standard ultrasound criteria for the disease were present: myometrial cysts, heterogenous myometrium, globular/asymmetric uterus, indistinct endo-myometrial junction and liner striations. Women not meeting these criteria served as controls. Women with fibroids, recurrent miscarriage and smokers were excluded. Implantation rates were calculated and logistic regression models were used to adjust for confounders for
Vol. 97, No. 3, Supplement, March 2012
P-65 Oocyte Defects Leading to Triploid Miscarriages After Donor Oocyte IVF. F.E. Marshall,a B. Behr,b R.B. Lathi.b a Department of Ob/Gyn, University of California Irvine Medical Center, Orange, CA; b Stanford Center for Reproductive Medicine, Stanford, Palo Alto, CA. BACKGROUND: The most common form of aneuploidy observed in first trimester miscarriages is trisomy, followed by polyploidy. Whereas, the risk of meiotic non-disjunction leading to trisomy increases with maternal age, the risk factors for triploid conceptions are poorly understood. Diandric triploidy is commonly thought to be due to polyspermia, but the etiology of and risk factors for digynic triploidy are unknown. OBJECTIVE(S): We present two cases of digynic triploidy after IVF with donor oocytes, leading to subsequent miscarriage. MATERIALS AND METHOD(S): Detailed chart reviewed two cases of triploid pregnancies after IVF cycle with donor oocytes. Both 26 year- old donors underwent controlled ovarian stimulation protocol using long Lupron with 75 IU rFSH and 75 IU hMG for 10 days. After adequate follicular response, 10,000 IU hCG was administered, and oocyte retrieval was performed 35 hours later. Oocytes were inseminated with approximately 25,000 motile sperm/ml and fertilization was evaluated 16-18 hours later. We evaluated IVF cycle parameters and sent miscarriage tissue for SNP microarray analysis. RESULT(S): Case 1: Peak E2 reached 4,375.7 and 30 oocytes retrieved. Post wash semen parameters: 41 M/ml with 91% motility and standard insemination performed. On day 1, three 2PNs were noted, four 3PN, and twenty-three non-fertilized. Embryologist noted poor oocyte quality and faint PNs. On day 3, the best two embryos were transferred: an 8 cell (grade I) from an embryo where no pronuclei were seen and a 6 cell (grade II) from a 2PN. Case 2: Peak E2 reached 3342, and 19 oocytes retrieved. Post wash semen parameters: 37 M/ml with 89% motility. On day 1, four 2PNs noted, and eleven non-fertilized. On day 3, the best two embryos were transferred: two 8 cell (grade II and compacting). The non-transferred embryos did not develop into blastocysts. Ultrasounds confirmed embryonic demise by 8 weeks. Chromosome analysis using SNP microarray revealed triploidy 69 XXX - maternal origin. CONCLUSION(S): These 2 cases illustrate that triploidy of maternal origin can cause miscarriage after oocyte donation. Our hypothesis is that low fertilization rate in the setting of normal semen parameters was a sign of poor oocyte quality. Therefore, we recommended to both couples that they consider a new oocyte donor for subsequent pregnancy attempts. Further studies are needed to evaluate the impact of patient characteristics, stimulation protocol, ovulation induction, and culture conditions on oocyte defects leading to triploidy.
P-66 Assisted Hatching of the Zona Pellucida Prior to Embryo Transfer in Fresh Donor Egg IVF Cycles. L. Cieslik, A. Khabani, C. Khabani, R. Dunsmoor-Su, J. Lamb, L. Hickok. Pacific NW Fertility, Seattle WA; University of Washington, Dept of Obstetrics and Gynecology, School of Medicine, Seattle WA.
S32
Abstracts
BACKGROUND: The early embryo is surrounded by the zona pellucida (ZP). Prior to implantation, the blastocyst must ‘‘hatch’’ out of the ZP so that implantation can occur. A variety of factors may influence the ability of the blastocyst to accomplish hatching and if so influence the chances for pregnancy occurring. Assisted hatching in in vitro fertilization cycles may improve the likelihood of implantation and pregnancy in certain age groups but it has not been well described in a donor/recipient IVF population. OBJECTIVE(S): The purpose of the study was to evaluate the effectiveness of day 5 assisted hatching of the zona pellucida prior to fresh embryo transfer in donor recipients. MATERIALS AND METHOD(S): This study was a retrospective casecontrol analysis of patients who underwent a fresh IVF cycle using donor eggs. The control group of 192 patients had embryos transferred prior to the implementation of assisted hatching of the ZP at our center in 2009 and 127 patients had an embryo transfer after assisted hatching of the ZP with CO2 laser during 2010. Cases and controls were compared using ttest and chi squared analyses. RESULT(S): There were no significant differences in age (41.5 vs 41.4 years), race, estradiol levels of the donor at the time of hCG administration (4130pg/ml vs 3965pg/ml), number of oocytes retrieved (23.5 vs 23.9) or M2s undergoing itracytoplasmic sperm injection (ICSI) (18.3 vs 18.9), number of oocytes fertilized (15.0 vs 16.1), blastocyst formation (9.2 vs 9.5) or the number of embryos transferred (1.66 vs 1.6). There was no difference in the biochemical pregnancy rate (78.7% vs 79.7%) or the number of gestational sacs (1.17 vs1.14). There was a non-statistically significant trend toward an increased probability of observing a fetal heartbeat (78.7% vs 76.6%, P¼0.65) but a trend toward an increase in live birth (75.6% vs 69.8%, P¼0.25) in cases versus controls. (Table1)
TABLE 1.
Chemical pregnancy rate Number of gestational sacs Clinical pregnancy rate Live birth or continuing pregnancy
Hatched (n¼127)
Controls (n¼192)
P value
78.7% 1.17 78.7% 75.6%
79.7% 1.14 76.7% 69.8%
0.83 0.70 0.65 0.25
CONCLUSION(S): This study is the first to evaluate the use of assisted hatching in donor egg IVF cycles. Day 5 Laser-assisted hatching of the zona pellucida prior to embryo transfer in donor IVF cycles may improve clinical pregnancy and live birth rates. FINANCIAL SUPPORT: none
P-67 Does Adenomyosis Impair Implantation? A Cohort Study. Z. Khan, MBBS,a D.M. Breitkopf, MD,b M. Al Hilli, MBBCh,b J.R. Jensen, MD,a C.C. Coddington, MD,a E.A. Stewart, MDa a Division of Reproductive Endocrinology & Infertility, Department of Obstetrics & Gynecology, Mayo Clinic, Rochester, MN 55905; b Department of Obstetrics & Gynecology, Mayo Clinic, Rochester, MN 55905. OBJECTIVE(S): The clinical impact of adenomyosis on infertility is controversial. The aim of this study was to determine the effect of ultrasound-diagnosed adenomyosis on implantation rates with good quality embryos in women undergoing IVF. MATERIALS AND METHOD(S): With IRB approval, all women undergoing fresh and frozen embryo transfers with good quality embryos (embryos at appropriate stage of development with minimal fragmentation) at Mayo Clinic, Rochester from January 2009 to December 2010 were included. Images from pre-IVF ultrasounds performed by the same ultrasonographer were reviewed independently by two physicians blinded to cycle outcome. Adenomyosis was diagnosed only when at least three of the five standard ultrasound criteria for the disease were present: myometrial cysts, heterogenous myometrium, globular/asymmetric uterus, indistinct endo-myometrial junction and liner striations. Women not meeting these criteria served as controls. Women with fibroids, recurrent miscarriage and smokers were excluded. Implantation rates were calculated and logistic regression models were used to adjust for confounders for
Vol. 97, No. 3, Supplement, March 2012