FISH Embryos Based on Gender

presented data collected from an Australian private reproductive medicine clinic. California recently approved a stem cell research initiative. Our results demonstrate for the first time that where public policy provides an opportunity for couples to donate their unused embryos to support stem cell research, this positively influences their decision on the fate of their unused embryos. (1) ACOG Committee on Ethics (2004), in Ethics in Obstetrics and Gynecology / American College of Obstetricians and Gynecologists 2nd Edition pp 37-41. (2) Catt J et al, (2003). Preimplantation Gender Selection, Proceedings of Fertility 2003, June 11-17, Aberdeen Scotland. Supported by: None P-494

on embryo morphology. These results may explain why certain egg donors have poor pregnancy conversion rates despite satisfactory ovarian response, oocyte quantity, and embryo morphologic appearance. In that case, PGD assessment of embryos created from donor eggs may improve implantation and conception rates, and minimize high order pregnancy rates through selection and transfer of only chromosomally competent embryos. However, more data on the genetic make up of the embryos of early reproductive age women is necessary in order to fully understand which factors such as spermatozoa, rate of mosaicism in embryos of young women, regime of ovarian stimulation or a consequence of an inherent error associated with the technique of single cell FISH contributes the most to the high rate of aneuploidy. Supported by: None P-493 Characteristics of Clients Seeking Gender Selection for Non-Medical Reasons. C. M. Briton-Jones, J. M. Steinberg. The Fertility Institutes, Encino, CA. OBJECTIVE: To identify and describe key characteristics and motivations of couples seeking gender selection for non-medical reasons. DESIGN: Prospective data collection from clients attending their first consultation for assisted reproductive treatment for gender selection. MATERIALS AND METHODS: 180 couples attending a consultation for assisted reproductive treatment for gender selection at a private fertility center in Los Angeles were included in this study. Same sex couples, couples choosing gender selection for medical reasons or who were primarily undertaking assisted reproductive treatment for infertility were excluded from the study. Routine questions asked in first patient consultations identified: any previous children and the gender of those children; the gender desired; the ethnicity of the individuals whether the male or the female partner initiated treatment investigation; the age of the female partner, the economic status of the couple and the preferred fate for embryos of the non-selected gender. The Chi Square statistical test for observed versus expected values was used. P⬍0.05 was considered significant. RESULTS: 99% of couples had one or more children only of the gender not requested. There was no significant bias for the selection of either gender for the subject group as a whole. However when divided into ethnic groups, there was a significant bias toward males in the Asian group where P⬍0.0001 and a significant bias towards females in the Caucasian group where P⬍0.001. There was no significant bias toward either the male or the female partner being the one who initiated investigation of the treatment. However, there was a highly significant correlation between the gender requested and the gender of the parent who initiated investigation of the treatment, where P⬍0.0001. 26% of couples predict that they will choose to discard the embryos of the non-desired gender, 72% predict that they will donate those embryos to research, particularly for embryonic stem cell research and 2% predict that they will donate embryos of the non-desired gender anonymously to an infertile couple. CONCLUSION: Overall there was no bias towards one gender. This is in conflict with what was published by the American College of Obstetrics and Gynecology (1) who predicted that offering gender selection for non-medical reasons “can be motivated by and reinforce the devaluation of women”. There was no significant bias towards one gender or the other with who was the first to initiate investigation of the treatment. However a strong correlation was found where the gender being sought and the gender of the partner initiating the treatment was the same. Our results support the findings of Catt, J et al (2) who

FERTILITY & STERILITY威

Comparison of Development Rates for PGD/FISH Embryos Based on Gender. S. L. Thormahlen, K. M. Delegge, T. F. O’Leary III, A. Finn, L. A. Scott, J. A. Hill. Fertility Centers of New England, Reading, MA. OBJECTIVE: Research in bovine and murine embryos suggests an increased development rate in male vs. female embryos. There has been some speculation that blastocyst ET weights gender due to this phenomenon but it has not been proved. With the introduction of IVF/PGD/FISH for aneuploidy screening, the ability to monitor human embryo development against gender can be realized. Because recent advances in PGD technologies have allowed fertility labs to look at the genetics, as well as the morphology, of embryos, this study also evaluates if embryo development in genetically abnormal embryos occurs at an increased rate to genetically normal embryos. DESIGN: A retrospective analysis of all PGD/FISH-Aneuploidy treatment embryos and their gender outcome from 2004, regardless of patient diagnosis. MATERIALS AND METHODS: 599 embryos were evaluated based on their euploidy: Group A-euploid embryos; Group B- monsomic embryos; Group C- trisomic embryos; and Group D- complex abnormal embryos. All embryos were cultured in sequential media and isolated into individual culture drops for assessment of development. Only viable embryos were included in this study. RESULTS: The table below shows no significant difference in gender outcome within all study groups. Female embryos were XX and male XY, only. Embryos with abnormal sex chromosomes were excluded.

118 blastocysts developed by day 5 of culture (20%). A blastocyst included early, full, and hatching blastocysts. Blastocyst development

S329

within the 4 study groups is shown in Table 2. There was no significant difference in development of blastocysts from male or female embryos or euploid (47%) vs. aneuploid embryos (53%; 63/118). CONCLUSION: Human male embryos do not appear to have a developmental advantage over female embryos in vitro, both having the same rate of blastocyst formation. However, euploid embryos develop to the blastocyst stage at an equal rate (467%) as aneuploid ones (53%; 63/118). This data corroborates previous published data that indicate blastocyst development does not eliminate abnormalities and suggests no gender skewing in either blastocyst culture or aneuploidy studies. Supported by: None

P-495 Is There a Relationship Between Early Embryo Cleavage and Genetic Abnormalities? Lessons From Preimplantation Genetic Diagnosis (PGD). S. Hamamah, V. Loup, T. Anahory, A. Girardet, B. Hedon, H. Dechaud. IVF Center, Montpellier, France; Molecular Biology, Montpellier, France; Ob-Gyn, Arnaud de Villeneuve Hospital, Montpellier, France. OBJECTIVE: Assessment of early human embryo cleavage (25-27 H) post-intracytoplasmic sperm injection (ICSI) has proven a reliable parameter for embryo viability selection with the highest capability of implantation and successful pregnancy. We have observed in our ICSI and PGD programs that the frequency of embryos reaching the 2-cell stage at 25-27 h post-ICSI is 22.0 % and 44.7 % respectively. The aim of the present study is to evaluate (i) the frequency of early cleavage embryos (25-27 H) in couples at risk of transmitting a monogenic disorder and chromosomal abnormalities; and (ii) the frequency of the early cleavage embryos in affected and uneffected embryons. DESIGN: 16 couples underwent 25 PGD cycles were included in this study. 18-19 h post-ICSI, all oocytes were evaluated for intactness and fertilization (2PN). 25-27 h post-ICSI, 185 embryos were checked for early cleavage (EC) or no early cleavage (NEC). MATERIALS AND METHODS: On day 2, the quality of embryos was assessed. According the number of anucleate fragments, the embryos were subdivided into grade A (no anucleate fragments), B (⬍ 20 % anucleate fragments, C (21 to 50 % anucleate fragments) and D (ⱖ50 % anucleate fragments) embryos. In the morning of day 3, grade A and B embryos were biopsied. Two blastomeres were removed from the embryos which contained ⱖ 6 blastomeres. Between the time of embryo biopsy and embryo transfer, embryos were kept in culture medium (G2.3). Only unaffected embryos (not carrying tested mutations or chromosomal abnormalities) were accepted for transfer. Available unaffected embryos were replaced on day 4. RESULTS: In 45 biopsied embryos (24 %), we could not obtain a diagnosis, while 69 embryos were affected by the disease under consideration (49 %) and 71 embryos (50 %) were non-affected. In 49 % (24/49) of unaffected embryos and 40 % (18/45) of affected embryos, EC embryos were detected at 25-27h post-ICSI. However, NEC embryos were detected in 51 % (25/49) of unaffected and 60% (27/45) of affected embryos. Amongst non-affected embryos, those selected for transfer provided from 50% and 64% of EC and NEC embryos respectively. 6 pregnancies were obtained from unaffected embryos irrespective of early cleavage rate. CONCLUSION: This is the first report which shows the frequency of EC 25-27 h post-ICSI in PGD cycles for monogenic or chromosomal disorders than in regular ICSI cycles. The frequency of EC is lower in affected embryos than in non-affected embryos and the NEC of non-affected embryos and in the affected embryo group the frequency of NEC is significanlty higher than EC rate. The success rate of PGD for monogenic diseases and chromosomal abnormalities was not associated with the percentage of EC of non affected-embryos. Supported by: None

P-496 Preimplantation Genetic Diagnosis (PGD) as a Beneficial Tool in Women With Recurrent Pregnancy Loss (RPL) and Advanced Maternal Age (AMA). L. B. Werlin, A. Decherney, I. Rodi, M. Kettel, B. Shapiro, S. Munne. Coastal Fertility Medical Center, Irvine, CA; Genesis Network for Reproductive Health, Irvine, CA; Reprogenetics, San Francisco, CA.

S330

Abstracts

OBJECTIVE: To report an update of the ongoing IRB approved, randomized, prospective study looking at patients who are at higher risk of producing aneuploid embryos. In a previous phase of the trial it was demonstrated that PGD may be beneficial in the RPL group, but it was not clear whether PGD would be beneficial in the AMA group. In an effort to determine if PGD is beneficial in both groups, the study was expanded. DESIGN: IRB approved, randomized, prospective study. MATERIALS AND METHODS: A total of 76 patients were enrolled from Aug 1, 2001 - April 1, 2004. All patients were randomized into either controls or PGD. All undedrwent stimulation protocols using Follistim and Antagon (Organon, West Orange, NJ). At the appropriate time, Pregnyl (hCG) 10,000 IU (Organon) was administered. Approximately 34-36 hours after hCG adminstration, ultrasound guided oocyte retrieval was performed. Intracytoplasmic sperm injection was performed on all mature oocytes. In the PGD group, embryo biopsy and blastomere fixation were done on day 3 post retrieval on all 6 - 8 cell embryos, with only one cell biopsied per embryo in most cases. Slides were then sent to Reprogenetics for flourescence in situ hybridization analysis for chromosomes 13, 15, 16, 17, 18, 21, 22, X and Y. Results were received on day 4 post retrieval. Embryo transfer (ET) of only the chromosomally normal embryos was done on day 5 post retrieval. In the control group, ET was performed on day 3 or 5 post retrieval based on physician preference. In both the PGD and control groups, corticosteroids, low-dose asprin (80-81 mg), and progestational supplementation were used. Serum B-hCG levels were obtained 12 days after ET. Pregnancy was confirmed by demonstrating at least two appropriate consecutively rising B-hCG levels. RESULTS: See table.

CONCLUSION: 1.) It was reconfirmed that in both groups of patients, PGD may be beneficial since there is a tendency for higher pregnancy rate, a clear higher implantation rate, and lower number of embryos replaced in the PGD group. 2.) If similar number of embryos were to be replaced in both groups, it would appear that PGD would be superior. 3.) In contrast, in European studies in which two cells per embryo were biopsied, PGD did not show any improvement over the control group. Some studies have indicated a significant reduction of implantation potential after 2-cell biopsy, explaining the European results. Supported by: Organon Pharmaceuticals, Inc., West Orange, NJ P-497 Sex Ratio and Cleavage Rhythm in Embryos From ICSI. L. M. Farah, L. M. Rossi, T. C. Bonetti, R. Joffe, A. Iaconelli Jr., E. Borges Jr.. Fertility - Assisted Fertilization Center, Sao Paulo, Brazil. OBJECTIVE: There are some controversies related to the prevalence of male and female embryos after assisted reproductive techniques. This ratio called primary sex ratio is, in fact, the rate between XX and XY embryos at the time of fertilization. In accordance to Pergament et al in 2002, in humans this ratio differs remarkably from the theoretically expected equality of 1: 1, and may be as high as 170 XY (males) to 100 XX (females). Moreover, it has been supposed that the injection of a single sperm directly into the egg appears to slow down the growth rate of the resulting female (but not male) early embryos. The aims of this study had been access the rate between embryos XX and XY from ICSI cycles and evaluate their cleavage rhythm and development.

Vol. 84, Suppl 1, September 2005