Elevated aneuploidy rates in embryos derived from immature oocytes from controlled ovarian hyperstimulation (COH) IVF cycles

of total fertilization failure with conventional insemination. These data are important for counseling patients regarding the outcomes of fertilization during IVF. Supported by: None. P-688 BIRTH FOLLOWING A SINGLE VITRIFIED- WARMED BLASTOCYST TRANSFER AFTER ICSI USING ROUND HEADED SPERM AND ASSISTED OOCYTE ACTIVATION IN A GLOBOZOOSPERMIC PATIENT. H. Hattori, C. Nishinaka, Y. Nakajo, H. Abe, Y. Araki, K. Kyono. Kyono ART Clinic, Sendai, Miyagi, Japan; Grad. Prog. of Human Sensing and Functional Sensor Engineering, Yonezawa, Yamagata, Japan; The Institute for ARMT, Gunma, Japan. OBJECTIVE: Birth following the transfer of a single vitrified-warmed blastocyst(VWB) after ICSI and assisted oocyte activation(OA)in a globozoospermic infertile man(GIM). DESIGN: A case report. MATERIALS AND METHODS: A 29-year-old woman and her 30-yearold husband who failed repeatedly following IUI and IVF, came to us with primary infertility of 2 years’ duration. The husband’s semen analysis was volume;3.0ml, density;18x106/ml, and motility;13.3%. All sperm showed round head(RH) shape morphologically. Mouse OA was tested with SrCl2. Sperm morphological figures were observed and processed for transmission electron microscopy (TEM). Ovarian stimulation with GnRH agonist-long protocol was started. After retrieval of oocytes and ICSI, assisted OA with calcium ionophore A23187(CI) was performed. All fertilized embryos were cultured to blastocyst formation, and blastocysts were frozen by using vitrification method. After two months, one VWB was transferred to the woman during another cycle which stimulated hormone replacement treatment. RESULTS: When RH sperm were injected into mouse oocytes, no oocytes were observed to have 2PN; however, 100% fertilization (2PN) was achieved by SrCl2 OA after ICSI. Sperm were observed to be 100% RH with abnormal morphology on light and TEM. All sperm heads were missing from acrosome region to the tip of the heads. Therefore, it is noted that these sperm really have defective OA substance. In this case, artificial activation was necessary to ensure pregnancy. Twenty-one oocytes were retrieved, and RH sperm were injected into 17 oocytes in metaphase II. Assisted OA with CI after ICSI resulted in a high fertilization rate (15 of 17, 88.2%) and developed 10 blastocysts on day 5 and one blastocyst on day 6 after insemination. All 11 blastocysts were cryopreserved by vitrification. After 2 months, one VWB (Gardner’s criteria 4BB; day 5) was transferred. As clinical pregnancy was recognized by observation of fetal heart beats in the gestational sac by ultra-sonography. The patient delivered a healthy male baby weighing 3180g at 40 weeks gestation by cesarean section on April 12, 2008. Apgar score was 9 at one minute, and 10 at 5 minutes. There were no congenital abnormalities. CONCLUSIONS: We report a successful birth from a single VWB after ICSI and assisted OA via CI in a GIM. To the best of our knowledge, this report of a birth using both ICSI with OA and a single VWB transfer is the first case in the world. Supported by: None. P-689 ELEVATED ANEUPLOIDY RATES IN EMBRYOS DERIVED FROM IMMATURE OOCYTES FROM CONTROLLED OVARIAN HYPERSTIMULATION (COH) IVF CYCLES. N. L. Buehler, C. Briton-Jones, M. Surrey, H. Danzer, D. L. Hill. Embryology, ART Reproductive Center, Beverly Hills, CA; Southern California Reproductive Center, Beverly HIlls, CA; Clinical, Southern California Reproductive Center, Beverly Hills, CA. OBJECTIVE: To evaluate the fertilization, embryo development and ploidy of embryos derived from metaphase I oocytes obtained from IVF cycles. DESIGN: Case Cohort Review. MATERIALS AND METHODS: Oocytes were retrieved 34-36 hours post hCG injection, oocytes were denuded with hyaluronidase immediately after oocyte retrieval. Intracytoplasmic Sperm Injection (ICSI) was performed 3940 hours post hCG injection. Resulting embryos were cultured using Vitrolife G-series Plus medium. Embryo quality was assessed on the morning of Day 3 (D3) then again on the morning of D5. Embryo biopsy was performed using a Zilos laser for zona opening to facilitate removal of blastomeres in Ca/Mg free media, and fixation of blastomeres performed using a combination of

FERTILITY & STERILITYÒ

hypotonic citrate solution followed by Carnoy’s fixative. FISH analysis was performed for chromosomal aneuploidy of chromosomes 8,13,14,15,16,17,18,20,21,22,X and Y. RESULTS: 302 embryos were derived from eggs assessed to be at the stage of metaphase I following denudation, but subsequently converted to the metaphase II stage prior to the ICSI procedure. 119 (39%) of these oocytes with delayed maturity fertilized with 2pn, 5(2%) fertilized with 1 pronucleus, 156(52%) did not produce pronuclei, and 18(6%) were not viable at fertilization assessment. 36(30%) of the zygotes displaying normal fertilization reached 6 or more cells on day 3 and 7(19%) of these created good quality blastocysts. 26(34%) of the embryos were screened for aneuploidy and 4(15%) were shown to be euploid for the chromosomes tested. CONCLUSIONS: Our data supports that of DeVos (et al, 1999) and Shu (et al, 2007), where immature oocytes derived from COH are capable of fertilization and good embryo development, but this occurs at a lower rate than expected for mature oocytes. Low fertilization rates, and high aneuploidy rates may be indicative of incompetence either inherent, or caused by maturation of these oocytes from metaphase I to metaphase II in vitro. Polar body biopsy may shed further light on these preliminary observations. Assessing the true clinical importance of immature oocytes is clear, particularly in cycles where few or no mature oocytes are obtained. The clinical competency of immature oocytes derived from COH is critical for appropriate counseling of patients who may be considering the transfer of embryos derived from such oocytes. Supported by: None.

P-690 SERUM ANTI-MULLERIAN HORMONE LEVELS IN NORMOOVULATORY AND ANOVULATORY WOMEN WITH POLYCYSTIC OVARIES: RELATIONSHIP TO THE OVARIAN FOLLICULAR ARREST. J. Qiao, J. Li, S. Zhao, X. Zhang. Reproductive Medical Center, Peking University Third Hospital, Beijing, China. OBJECTIVE: To compare serum and follicular fluid (FF) AMH levels in normoovulatory and anovulatory women with PCO and evaluate the relationship between AMH and the follicular arrest of the infertile women with PCO. DESIGN: Case-control study. MATERIALS AND METHODS: Thirty-five normoovulatory women with normal ovarian morphology (group A, control), 29 normoovulatory women with PCO (group B) and 38 anovulatory women with PCO (group C) underwent controlled ovarian hyperstimulation (COH) with the short protocol. Serum AMH levers on the day 0 and 6 of gonadotropins treatment (d0 or d6) and the day of HCG administration (dHCG) were measured using enzyme-linked immunosorbent assay (ELISA). FF AMH levels on the day of oocyte pickup (dOPU) were measured using same method as serum assay. RESULTS: No significant difference in serum AMH levels of d0, d6 and dHCG during COH was found between group B and C (5.87 vs. 7.99, 3.92 vs. 5.54, and 2.45 vs. 2.90ng/ml, respectively; P>0.05), but significantly higher than group A. (2.52, 1.17, and 0.79ng/ml, respectively; P<0.01). FF AMH levels have no difference between group B and C (54.77 vs. 64.05ng/g protein; P>0.05), but significantly higher than group A (29.45 ng/g protein; P<0.05). Significant correlation was found between serum AMH (d0, d6 or dHCG) and follicle numbers (d0 or dOPU). TABLE 1. Serum and FF AMH levels during COH (median and range) time serum AMH d0 levels (ng/ml) d8 dHCG FF AMH levels dOPU (ng/g protein)

Group A (n¼35)

Group B (n¼29)

Group C (n¼38)

2.52 (0.22-10.80) 5.87 (2.34-15.60)b 7.99 (3.13-14.80)a 1.17 (0.10-5.53) 3.92 (1.14-13.10)a 5.54 (1.84-11.70)a 0.79 (0.03-2.88) 2.45 (0.67-9.23)a 2.90 (1.13-7.00)a 29.45 (8.1-160.9) 54.77 (22.3-203.3)c 64.05 (19.6-281)b

Group A¼ normoovulatory women with normal ovarian morphology; Group B¼ normoovulatory women with PCO;Group C¼ anovulatory women with PCO. a compared with group A, P<0.001. b compared with group A, P<0.01. c compared with group A, P<0.05.

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