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The optimal timing of blastocyst vitrification after trophectoderm biopsy of preimplantation genetic screening
P-73 Tuesday, October 18, 2016 A NOVEL PREDICTIVE MODEL OF THE PROBABILITY OF LIVE BIRTH WHEN USING VITRIFIED HUMAN OOCYTES AS A MEANS OF FERTILITY PRESERVATION. A. L. Mauner,a E. Surrey,b R. L. Gustofson,b L. A. Kondapalli,b W. B. Schoolcraft.b aEmbryology, Fertility Laboratories of Colorado, Lone Tree, CO; bColorado Center for Reproductive Medicine, Lone Tree, CO. OBJECTIVE: Develop a predictive model to aid physicians in providing patients a realistic probability of live birth in consideration of their post oocyte retrieval data. DESIGN: Cross-sectional Analysis. MATERIALS AND METHODS: A retrospective cross-sectional analysis of patients who previously vitrified oocytes in an attempt to conceive at the Colorado Center for Reproductive Medicine was conducted. A sample of 137 patients who thawed oocytes from 2007 to 2015 which had data for all relevant predictor variables and whose oocytes were originally cryopreserved using the Kuwayama Cryotop Vitrification method were used for analysis. Both demographic and clinical predictors were used to develop a prediction model using a supervised forward selection approach. Statistical analysis was conducted using SAS 9.4 and R 3.2.3. RESULTS: The study population consisted of patients aged 19-44 (mean std. error: 27.27 0.40) with average Anti-M€ullerian hormone (AMH) levels of 4.00 0.20, day 3 Follicle Stimulating hormone (FSH) levels of 6.83 0.47, Body Mass Index (BMI) of 24.21 0.32, warming 15.98 0.44 oocytes, and transferring 1.74 0.04 embryos. The final model contained number of oocytes warmed, AMH level of oocyte source, percentage of post retrieval oocytes that were mature, age of oocyte source, total gonadotropin dosage, day 3 FSH level of oocyte source, number of days oocytes were cryopreserved, number of days on stimulation medication and BMI of oocyte source. This model had a C index of 0.779 (bootstrapped 95% CI 0.676, 0.821), a Brier score of 0.190 (bootstrapped 95% CI 0.176, 0.221), and a Hosmer-Lemeshow test statistic of 0.646. CONCLUSIONS: Overall calibration of the model was good, except for upper bound probabilities (60-100%) where the model slightly over-estimated probabilities. While acknowledging its limitations, this model will be translated into an application for physician use. This will serve as an invaluable tool for physicians as they consult with patients on their future probability of success and the potential for requiring multiple oocyte vitrification cycles to achieve a live birth.
CRYOPRESERVATION AND FROZEN EMBRYO TRANSFER P-74 Tuesday, October 18, 2016 THE OPTIMAL TIMING OF BLASTOCYST VITRIFICATION AFTER TROPHECTODERM BIOPSY OF PREIMPLANTATION GENETIC SCREENING. C. Lee,a,b T. Lee,a,c M. Lee.a,c aDepartment of Obstetrics and Gynecology, Chung Shan Medical University Hospital, Taichung, Taiwan; bDepartment of Obstetrics and Gynecology, National Taiwan University Hospital, Taipei, Taiwan; cInstitute of Medicine, Chung Shan Medical University, Taichung, Taiwan. OBJECTIVE: Vitrification was recommended for cryopreservation of biopsied blastocysts. We investigated whether the timing of vitrification after trophectoderm (TE) biopsy for preimplantation genetic screening (PGS) program associated with the success of implantation and pregnancy after frozen blastocyst transfer. DESIGN: A retrospective cohort study. MATERIALS AND METHODS: There were 1,329 blastocysts from 223 patients underwent the TE biopsy and array CGH. The PGS and frozen blastocyst transfer cycles were perfromed from Dec, 2012 to May, 2015. Only the good blastocyst (Gardner grading system: grade 4, 5 and 6) on day 5 or day 6 were selected for biopsy. All blastocyst were underwent vitrification immediately (0.5 hour) or 1 to 6 hours after biopsy. At the time of vitrification, the interval between TE biopsy and vitrification and the morphology of blastocyst re-expansion relative to original expansion cavity was recorded. The re-expansion grades were including (1) without significantly re-expansion (NE) (2) 3/4 re-expansion (3/4E) (3) full re-expansion or hatching. The euploidy blastocyst(s) (12 embryos) was/were selected first for transfer on the next cycle. According to the transferred blastocyst which showed different re-expansion grade at time of vitrification, all patients were divided into four groups: (Group I: non-expansion (n¼40), Group II: 3/4 re-expansion (n¼29) and Group III: all re-expansion or hatching (n¼154)).
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RESULTS: At 0.5 hour after biopsy, 100% of biopsied blastocyst was without significantly re-expansion; at 1 hour after biopsy, only 5.7% of biopsied blastocyst reached 3/4E or E grade; at 3 hours after biopsy, more than 86% blastocyst reached 3/4E or E. The survival rate of biopsied blastocyst was 100% (357/357) after warming. The implantation rate in Group III (63.1%, 159/252) was significantly higher than that in Group I (45.2%, 28/ 62; p¼0.010). The clinical pregnancy rates in Group III (73.4%, 113/154) and Group II (75.9%, 22/29) were significantly higher than that in Group I (52.5%, 21/40; p¼0.011 and p¼0.048, respectively). Combining two factors which including the re-expansion grades and timing of vitrification, the implantation and clinical pregnancy rates in the Group of biopsied blastocyst with R3/4E and intervalR3 hours (63.7%, 179/281; 74.0, 128/173, respectively) were significantly higher than that in the Group of biopsied blastocyst with NE and interval<3 hours (45.3%, 24/53; 50.0, 17/34; p¼0.012 and p¼0.005, respectively). The optimal timing of vitrification is biopsied blastocyst keeping culturing to reach R3/4 re-expansion and R3 hour after TE biopsy. CONCLUSIONS: The optimal timing of vitrification after TE biopsy provides high implantation and pregnancy rates after frozen blastocyst transfer. References: 1. Ahlstr€om A, Westin C, Wikland M, Hardarson T. Prediction of live birth in frozen-thawed single blastocyst transfer cycles by pre-freeze and post-thaw morphology. Hum Reprod 2013;28:1199-1209. 2. Goto S, Kadowaki T, Tanaka S, Hashimoto H, Kokeguchi S, Shiotani M. Prediction of pregnancy rate by blastocyst morphological score and age, based on 1,488 single frozen-thawed blastocyst transfer cycles. Fertil Steril 2011;95:948-952.
P-75 Tuesday, October 18, 2016 EFFECT OF COLONY-STIMULATING GROWTH FACTOR ON OUTCOME OF FROZEN-THAWED EMBRYO TRANSFER IN PATIENTS WITH REPEATED IMPLANTATION FAILURE. D. Obidniak,a A. Gzgzyan,b L. Dzhemlikhanova,a A. Feoktistov.c aMedical Faculty, Saint-Petersburg State University, Saint-Petersburg, Russian Federation; b Saint-Petersburg State University, Saint-Petersburg, Russian Federation; c Medical Faculty, Medical Group ‘‘Mother and Child’’, Saint-Petersburg, Russian Federation. OBJECTIVE: To investigate the granulocyte colony-stimulating growth factor (G-CSF) effect on frozen-thawed embryo transfer outcome depending on its route of administration in patients with repeated implantation failure (RIF). DESIGN: randomized controlled trial MATERIALS AND METHODS: After obtaining institutional review board approval 130 women aged 32 - 40 years were involved. Inclusion criteria were: RIF defined as at least two cycles of in vitro fertilization in which good-quality embryos (The Gardner blastocyst grading system) was transferred in each cycle without achieving a clinical pregnancy; the availability of embryos vitrified at the blastocyst stage. Exclusion criteria were: congenital uterine anomalies; asherman’s syndrome; endometrial thickness on the day of embryo transfer in previous cycles was less than 7 mm. 3 groups of patients were formed on the basis of randomization: 1) study group N1 (N ¼ 40): intrauterine perfusion with G-CSF (filgrastim 30 million IU, 1 mL) using insemination catheter 5 days prior to embryo transfer; 2) study group N2 (N ¼ 30): G-CSF (filgrastim 30 million IU, 1 ml) was administered subcutaneously once at the day of embryotransfer; 3) Control group: no therapy (N ¼ 60). Patients of all groups did not differ statistically on age, body mass index, number of unsuccessful attempts. Endometrium preparation was carried out according to standardized protocol of hormone replacement therapy with ultrasound monitoring: estradiol valerate 6 mg/day started on day 3 of menstrual period without prior pituitary down-regulation; the luteal phase support consisted of micronized progesterone vaginally in a daily dose of 800 mg. The transfer of viable embryos was performed on the 6-th day of progesterone administration. The treatment effectiveness was assessed by the clinical pregnancy rate and implantation rate. Clinical pregnancy was defined as ultrasound detection of fetal cardiac activity. Statistical significance of the observed effects was tested by P values and confidence intervals. RESULTS: The clinical pregnancy rate in the study group N1, 2 and the control group consisted 37.5% (15/40); 46.6% (14/30); 26.6% (16/60) respectively. The implantation rate differed significantly: it revealed 31.2% in the study group N1; in the study group N 2 - 38.6%; in the control group - 19.8%. No adverse event was noted. CONCLUSIONS: - G-CSF enhances the implantation rate and clinical pregnancy rate in frozen-thawed embryo transfer;
Vol. 106, No. 3, Supplement, September 2016
CRYOPRESERVATION AND FROZEN EMBRYO TRANSFER P-74 Tuesday, October 18, 2016 THE OPTIMAL TIMING OF BLASTOCYST VITRIFICATION AFTER TROPHECTODERM BIOPSY OF PREIMPLANTATION GENETIC SCREENING. C. Lee,a,b T. Lee,a,c M. Lee.a,c aDepartment of Obstetrics and Gynecology, Chung Shan Medical University Hospital, Taichung, Taiwan; bDepartment of Obstetrics and Gynecology, National Taiwan University Hospital, Taipei, Taiwan; cInstitute of Medicine, Chung Shan Medical University, Taichung, Taiwan. OBJECTIVE: Vitrification was recommended for cryopreservation of biopsied blastocysts. We investigated whether the timing of vitrification after trophectoderm (TE) biopsy for preimplantation genetic screening (PGS) program associated with the success of implantation and pregnancy after frozen blastocyst transfer. DESIGN: A retrospective cohort study. MATERIALS AND METHODS: There were 1,329 blastocysts from 223 patients underwent the TE biopsy and array CGH. The PGS and frozen blastocyst transfer cycles were perfromed from Dec, 2012 to May, 2015. Only the good blastocyst (Gardner grading system: grade 4, 5 and 6) on day 5 or day 6 were selected for biopsy. All blastocyst were underwent vitrification immediately (0.5 hour) or 1 to 6 hours after biopsy. At the time of vitrification, the interval between TE biopsy and vitrification and the morphology of blastocyst re-expansion relative to original expansion cavity was recorded. The re-expansion grades were including (1) without significantly re-expansion (NE) (2) 3/4 re-expansion (3/4E) (3) full re-expansion or hatching. The euploidy blastocyst(s) (12 embryos) was/were selected first for transfer on the next cycle. According to the transferred blastocyst which showed different re-expansion grade at time of vitrification, all patients were divided into four groups: (Group I: non-expansion (n¼40), Group II: 3/4 re-expansion (n¼29) and Group III: all re-expansion or hatching (n¼154)).
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RESULTS: At 0.5 hour after biopsy, 100% of biopsied blastocyst was without significantly re-expansion; at 1 hour after biopsy, only 5.7% of biopsied blastocyst reached 3/4E or E grade; at 3 hours after biopsy, more than 86% blastocyst reached 3/4E or E. The survival rate of biopsied blastocyst was 100% (357/357) after warming. The implantation rate in Group III (63.1%, 159/252) was significantly higher than that in Group I (45.2%, 28/ 62; p¼0.010). The clinical pregnancy rates in Group III (73.4%, 113/154) and Group II (75.9%, 22/29) were significantly higher than that in Group I (52.5%, 21/40; p¼0.011 and p¼0.048, respectively). Combining two factors which including the re-expansion grades and timing of vitrification, the implantation and clinical pregnancy rates in the Group of biopsied blastocyst with R3/4E and intervalR3 hours (63.7%, 179/281; 74.0, 128/173, respectively) were significantly higher than that in the Group of biopsied blastocyst with NE and interval<3 hours (45.3%, 24/53; 50.0, 17/34; p¼0.012 and p¼0.005, respectively). The optimal timing of vitrification is biopsied blastocyst keeping culturing to reach R3/4 re-expansion and R3 hour after TE biopsy. CONCLUSIONS: The optimal timing of vitrification after TE biopsy provides high implantation and pregnancy rates after frozen blastocyst transfer. References: 1. Ahlstr€om A, Westin C, Wikland M, Hardarson T. Prediction of live birth in frozen-thawed single blastocyst transfer cycles by pre-freeze and post-thaw morphology. Hum Reprod 2013;28:1199-1209. 2. Goto S, Kadowaki T, Tanaka S, Hashimoto H, Kokeguchi S, Shiotani M. Prediction of pregnancy rate by blastocyst morphological score and age, based on 1,488 single frozen-thawed blastocyst transfer cycles. Fertil Steril 2011;95:948-952.
P-75 Tuesday, October 18, 2016 EFFECT OF COLONY-STIMULATING GROWTH FACTOR ON OUTCOME OF FROZEN-THAWED EMBRYO TRANSFER IN PATIENTS WITH REPEATED IMPLANTATION FAILURE. D. Obidniak,a A. Gzgzyan,b L. Dzhemlikhanova,a A. Feoktistov.c aMedical Faculty, Saint-Petersburg State University, Saint-Petersburg, Russian Federation; b Saint-Petersburg State University, Saint-Petersburg, Russian Federation; c Medical Faculty, Medical Group ‘‘Mother and Child’’, Saint-Petersburg, Russian Federation. OBJECTIVE: To investigate the granulocyte colony-stimulating growth factor (G-CSF) effect on frozen-thawed embryo transfer outcome depending on its route of administration in patients with repeated implantation failure (RIF). DESIGN: randomized controlled trial MATERIALS AND METHODS: After obtaining institutional review board approval 130 women aged 32 - 40 years were involved. Inclusion criteria were: RIF defined as at least two cycles of in vitro fertilization in which good-quality embryos (The Gardner blastocyst grading system) was transferred in each cycle without achieving a clinical pregnancy; the availability of embryos vitrified at the blastocyst stage. Exclusion criteria were: congenital uterine anomalies; asherman’s syndrome; endometrial thickness on the day of embryo transfer in previous cycles was less than 7 mm. 3 groups of patients were formed on the basis of randomization: 1) study group N1 (N ¼ 40): intrauterine perfusion with G-CSF (filgrastim 30 million IU, 1 mL) using insemination catheter 5 days prior to embryo transfer; 2) study group N2 (N ¼ 30): G-CSF (filgrastim 30 million IU, 1 ml) was administered subcutaneously once at the day of embryotransfer; 3) Control group: no therapy (N ¼ 60). Patients of all groups did not differ statistically on age, body mass index, number of unsuccessful attempts. Endometrium preparation was carried out according to standardized protocol of hormone replacement therapy with ultrasound monitoring: estradiol valerate 6 mg/day started on day 3 of menstrual period without prior pituitary down-regulation; the luteal phase support consisted of micronized progesterone vaginally in a daily dose of 800 mg. The transfer of viable embryos was performed on the 6-th day of progesterone administration. The treatment effectiveness was assessed by the clinical pregnancy rate and implantation rate. Clinical pregnancy was defined as ultrasound detection of fetal cardiac activity. Statistical significance of the observed effects was tested by P values and confidence intervals. RESULTS: The clinical pregnancy rate in the study group N1, 2 and the control group consisted 37.5% (15/40); 46.6% (14/30); 26.6% (16/60) respectively. The implantation rate differed significantly: it revealed 31.2% in the study group N1; in the study group N 2 - 38.6%; in the control group - 19.8%. No adverse event was noted. CONCLUSIONS: - G-CSF enhances the implantation rate and clinical pregnancy rate in frozen-thawed embryo transfer;
Vol. 106, No. 3, Supplement, September 2016