P-74 APPLICATION OF HUMAN UMBILICAL CORD SERUM (hUS) IN IN VITRO MATURATION (IVM) MEDIUM AND ESTABLISHMENT OF CLINICAL PREGNANCIES. Z. Zhiguo, C. Yunxia. Medical Repro- ductive Center, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China. OBJECTIVE: To explore a new protocol in IVM by using a modified IVM medium containing hUS aiming at improving clinical pregnancy rate and the level of IVM technology. DESIGN: Prospective cohort study. MATERIALS AND METHODS: Immature oocytes from 47 cycles of 47 patients with polycystic ovary syndrome (PCOS) were respectively placed into IVM I medium (containing hFF, hFF group) and IVM II medium (con- taining hUS, hUS group) for in vitro maturation. Finally, effect of hUS on the outcome of IVM were discussed and analyzed. Furthermore, effect of culture time on mature rate was explored and analyzed in the study. RESULTS: In hUS group (15 cycles), a total of 160 immature oocytes were obtained. During the process of IVM, 93.75% of maturation rate, 96.15% of fertilization rate, 96.0% of cleavage rate, 50% of high-quality em- bryo rate and 40% of clinical pregnancy rate were obtained. In hFF group (32 cycles), maturation rate, fertilization rate, cleavage rate, high-quality embryo rate and clinical pregnancy rate were 77.08%, 92.37%, 96.96%, 23.77% and 22.58% in sequence. According to the above outcome, maturation rate, fer- tilization rate, high-quality embryo rate and clinical pregnancy rate in hUS group were significantly higher than those in hFF group. Comparing matura- tion rate and high-quality embryo rate, respectively a significant difference was found between groups (P<0.05). CONCLUSIONS: IVM medium containing hUS appears to be much ef- fective to stimulate maturation of immature oocytes and capable of achieving a promising clinical outcome in IVM. Furthermore, it is vital to select an op- timal in vitro maturation time for immature oocyte in terms of different IVM medium. Supported by: The authors acknowledge the fund support from Anhui Province Education Bureau Project (grant number 2003kj213). The authors are also grateful to Doctor Xu Xiaofeng and Fu Xingchun for their kind help and technical assistance. P-75 POINT MUTATIONS IN OOCYTE MITOCHONDRIAL DNA (mtDNA) AND THEIR RELATIONSHIP TO CHANGES IN MITO- CHONDRIAL MEMBRANE POTENTIAL (DYm), MATERNAL AGE, INFERTILITY DIAGNOSIS AND IVF CYCLE OUTCOME. R. A. Forman, B. Acton, A. Jurisicova, R. F. Casper. Obstetrics and Gynecol- ogy, Mount Sinai Hospital/Samuel Lunenfeld Research Institute, Toronto, ON, Canada. OBJECTIVE: To determine if point mutations in oocyte mitochondrial DNA (mtDNA) located within the NADH dehydrogenase subunit 2 (S2) and cytochrome b (CYTO) are associated with changes in mitochondrial membrane potential (DYm), maternal age, infertility diagnosis and cycle outcome. DESIGN: Retrospective. MATERIALS AND METHODS: 56 metaphase II unfertilized oocytes were obtained from patients undergoing ART treatment. Oocytes were di- vided into groups based on maternal age. The membrane potential was de- termined on live cells using DYm sensitive JC-1 then stored at 80  C. The mutational survey in the oocytes was conducted by amplification of mtDNA with primers designed to span an area of known point mutations previously identified in individuals with heritable late-onset disorders such as essential hypertension and type II diabetes. Amplified products were sequenced from both ends and compared to a reference human mtDNA sequence. RESULTS: 56 metaphase II oocytes were chosen as a representative sam- ple from patients undergoing ART treatment. Oocytes were divided into groups based on maternal age. In the majority of cases there is one oocyte per patient. 56 oocytes underwent PCR amplification. Successful amplifica- tion of at least one of the two studied amplicons occurred in at least 50% of the samples. 24 amplicons were generated from the primer encompassing the S2 region. Six point polymorphisms/mutations were identified in this area. The most frequent polymorphism was located at position 4769 and occurred in 91.7% of amplicons. One oocyte with a point mutation at position 4917 and known to be associated with Lebner’s Hereditary Optic Neuropathy (LHON) was detected. 26 amplicons were generated from the primer encom- passing the CYTO region. Six point polymorphisms/mutations were identi- fied. 96% of oocytes contained an identifiable polymorphism. The most frequent one was at position 15326 and leads to a known amino acid change found in individuals or their families with type II diabetes. A novel point mutation was found at position 15497. All identified mtDNA mutations were in patients %35 years. Oocytes of patients with mutations in CYTO and S2 showed lower DYm values. CONCLUSIONS: One novel mtDNA mutation was identified in CYTO at position 15497. Five mtDNA mutations were identified (3 in CYTO and 2 in NADH S2). All identified mtDNA mutations were in patients %35 years. Oocytes of patients with mutations in CYTO and S2 showed lower DYm values. Supported by: None. P-76 INDIVIDUALIZED ASSESSMENT OF OOCYTE CUMULUS CELL HEALTH AND OOCYTE QUALITY, FERTILIZATION, EMBRYO DEVELOPMENT AND IMPLANTATION. P. A. Hassun, M. Ribeiro, A. M. Rocha, G. D. Smith, E. Motta, P. Serafini. Huntington Reproductive Medicine, Sao Paulo, SP, Brazil; Department of Ob/Gyn, Urology, and Physiology; Reproductive Sciences Program, University of Michigan, Ann Harbor, MI. OBJECTIVE: Predicting oocyte embryonic developmental and/or implan- tation potential has enormous clinical implications. However, non-invasive means of oocyte assessment are wanting. Cumulus cells (CCs) play an impor- tant role in follicle/oocyte cross-talk. Objectives were to characterize CC health in relation to individual oocytes using fluorescent activated cell sorting (FACS); and determine if percentage of viable, mid-apoptotic, or dead CCs were related to, or predictive of, oocyte quality, embryo development, and implantation rate (IR). DESIGN: Prospective laboratory study. MATERIALS AND METHODS: Oocyte cumulus complexes (OCCs; 238) were retrieved from 35 women age 37  0.8 (mean  SE) during con- trolled ovarian stimulation for ICSI treatment. Cumulus cells were washed, dispersed, and assessed as a group for each oocyte by FACS for viability, mid-apoptotic, and death using Guava ViaCount Reagents and flow cytome- ter. Oocyte quality (maturation, granualization, vacuolization, perivitelin space), fertilization, embryo cell number, embryo fragmentation, and IR were recorded. Nonparametric, parametric, correlation analyses, and predic- tive values were preformed with Chi-square, unpair Student’s t-test, Pear- son’s correlation coefficients, and linear regression. RESULTS: Percent viable (25% vs. 25%), mid-apoptotic (15% vs. 12%), and dead (60% vs. 63%) CCs were not significantly different between oocytes of normal and abnormal morphology, respectively. No significant correlation existed between oocyte maturation and percent viable, mid-ap- optotic or dead CCs. Of 215 oocytes injected 62% fertilized and percent viable (25% vs. 25%), mid-apoptotic (14% vs. 13%), and dead (61% vs. 62%) CCs were similar between oocytes that fertilized or did not, respec- tively. Percent viable, mid-apoptotic, and dead CCs did not change as a function of embryo development or degrees of fragmentation on day 2 or 3. Percent mid-apoptotic CCs were positively correlated with day 3 em- bryo blastomere number (P<0.02). Lastly, percent mid-apoptotic and dead CCs were positively and negatively correlated with implantation rate, re- spectively (P<0.01). CONCLUSIONS: Cumulus cell FACS can be performed for individual oocytes. A 1% increase in mid-apoptotic CCs predicted a 2.1% increate in IR and a 1% decrease in dead CCs predicted at 0.6% increase in IR. These findings suggest importance of assessing more than CC viability; that apopto- sis and death are import differential measures; and future studies should focus on intermediate markers of CC apoptosis and IR. Supported by: None. P-77 UNEXPECTED GENDER BIAS FOUND IN IVF CYCLES FOR SEX SELECTION. T. Mukherjee, M. Luna, A. De Venuta, E. Flisser, A. B. Copperman, J. Barritt. Reproductive Endocrinology and Infertility, S134 Abstracts Vol. 88, Suppl 1, September 2007