- Email: [email protected]
Blastomere symmetry is an important predictor of blastocyst development
either the dry or humid systems. Moreover after obtaining informed consent 32 4-8 cell human embryos were also randomly assigned to culture with one of the above systems. All media droplets were covered with mineral oil and the media were exchanged every 48 hours. Blastocyst development rates were evaluated after 3 days. RESULTS: For mouse embryos 93.1% and 91.9% developed to blastocyst stage with the dry and the humidfied incubator system, respectively. Similarly the blastocyst formation rates of human embryos used in the experiment were not significantly different 62.5% and 56.3% with the dry and humid incubator, respectively. CONCLUSION: Our results suggest that the development of both mouse and human embryos in a dry culture system were effectively identical to those of embryos cultured under standard humidified culture conditions. Although results are still preliminary regarding human application in our opinion, it might not be necessary to use a humidified atmosphere for culturing mouse or human embryos. The dry culture system used in the present study is efficient, hygienic and requires only little maintenance. Hence this new dry incubation system could reduce the risk of a possible contamination and contribute to lowering maintenance costs of an in-vitro fertilization program.
P-485 Wednesday, October 19, 2011 EFFECT OF EMBRYO GROUP CULTURE STRATEGY ON THE BLASTOCYST DEVELOPMENT AND PREGNANCY OUTCOME. T. Tao, A. Robichaud, J. Mercier, R. Ouellette. Conceptia Clinic, Moncton, NB, Canada. OBJECTIVE: The aim of this study is to investigate the influence of different embryo group culture strategy on the blastocyst development and pregnancy outcome. DESIGN: Two embryo group culture methods were compared. MATERIALS AND METHODS: Oocytes were inseminated either by standard IVF or ICSI. All zygotes were individually cultured in 20 ml droplets of G1.3 medium under oil at 37 C, in a humidified atmosphere containing 6% CO2, 5% O2 and 89 % N2 until day 3. On this day embryos were classified into two categories: good quality (have even, regular and spherical blastomeres, with R6 cells and %15% fragmentation) and poor quality. Embryos were then cultured in group of 2-5 embryos per droplet (50 ml G2.3) until day 5 or 6. The group culture strategy is: A, embryos were randomly grouped regardless of embryo quality; B, good or poor quality of embryos was separately grouped. Maximum of two blastocysts were transferred to patient on day 5. RESULTS: Results were shown in table 1. The blastocyst rate (62%) in group B were significantly higher than that in group A (43%). There was no statistical difference between two groups in the pregnancy rate. TABLE 1. Outcome of blastocyst development and pregnancy
Group # Patient A B
36 31
# embryos Average embryos cultured # Blastocysts transferred # Pregnancy 397 291
172 (43%) a 179 (62%) b
1.8 1.9
26 (72%) 20 (65%)
In group B, the blastocyst rates for good or poor quality of day 3 embryos were, respectively, 74% (159/215) and 26% (20/76), the average was 62%. a,bValues with different superscript letters within the same column are significant different (P<0.05). CONCLUSION: Our results suggest that day 3 embryos with good or poor quality should be separately grouped for culture to blastocyst stage. Autocrine or paracrine growth or survival factors produced by the embryos may support itself and other embryos in their development. Some other unknown factors released from poor quality embryos (such as arrested embryos) may inhibit the surrounding embryo development.
P-486 Wednesday, October 19, 2011 METABOLOMIC PROFILE OF EMBRYO CULTURE MEDIA UNDER HYPOXIA. J. M. De los Santos, P. Gamiz, F. Domınguez, A. Galan, S. Perez, M. J. De los Santos. IVF Laboratory, IVI Valencia, INCLIVA, Valencia, Spain; Embryomics S.L., Derio, Vizcaya, Spain. OBJECTIVE: Aim of this study was to determine whether human pre-embryos cultured under different oxygen concentrations (5% vs 20%) suffer from some sort of change in their metabolism.
S250
Abstracts
DESIGN: Prospective study. MATERIALS AND METHODS: A total of 23 patients of our ovum donation program were incluided in this study. 41 samples of spent embryo culture media (n ¼ 24; 5% [O2]; n ¼ 17; 20% [O2]) were used. Both groups selected had 100% of implantation rate. Embryos were cultured in O2/CO2 incubator. Culture media and controls were collected on day +3 and stored at -186 C. There were no differences on the age of the donors (26.2 vs 25.8), neither on the embryo quality. Samples were thawed and metabolites were extracted, then injected into ultra performance liquid chromatograph coupled to a mass spectrometer. All endogenous metabolites present in samples were measured, defined by its relation mass/charge and their retention time. For statistical analysis, univariate analysis (comparison of means) of each metabolite, between both groups and multivariate (principal component analysis) to find overall differences between groups were made. P values <0.05 were considered significant. RESULTS: Multivariate analysis of the global metabolomic profile of embryos cultured in hypoxia vs atmospheric conditions did not reveal significant differences between both groups. Despite no differences were observed in global profile, through univariate analysis, two metabolites defined by their relationship mass/charge and retention time, appeared significantly ‘‘high’’, while a third was diminished, in the group of embryos grown under low oxygen tensions. CONCLUSION: Culture of human pre-embryos in low oxygen tension did not change significantly their metabolic glycolysis pathways during in vitro development because there were no changes in the consumption profile of glucose, pyruvate or lactate. Furthermore, amino acids used as implantation markers such as Asn, Gly or Leu are not altered under both oxygen culture conditions.
P-487 Wednesday, October 19, 2011 BLASTOMERE SYMMETRY IS AN IMPORTANT PREDICTOR OF BLASTOCYST DEVELOPMENT. K. Nakayama, N. Fukunaga, R. Nagai, H. Kitasaka, Y. Hashiba, Y. Asada. Asada Ladies Clinic, Nagoya, Aichi, Japan; The Asada Institute for Reproductive Medicine, Kasugai, Aichi, Japan. OBJECTIVE: The aim of this study was to evaluate whether the blastomere symmetry of day 3 embryos affects blastocyst development on day 3. DESIGN: Retrospective study. MATERIALS AND METHODS: We reviewed the outcome of Day 5 embryo transfer (ET) using fresh embryos (436 cycles) and frozen and thawed embryos (489 cycles) between January 1, 2008 and December 31, 2009. Blastocyst development was assessed for embryos divided into two groups based on morphology: symmetrical group (uniform blastomeres) and unsymmetrical group (uneven blastomeres). All embryos had 6-8 cells and %20% fragmentation on Day 3. Chi-square test was used to examine the differences between the groups. RESULTS: Blastosyst development was significantly higher for the symmetrical group than the unsymmetrical group at all score levels for ET performed with fresh and frozen and thawed embryos (Table 1).
TABLE 1. Results of Day 5 ET for fresh and frozen and thawed embryos
Gardner’s score
fresh ET Symmetrical group Unsymmetrical group P frozen-thawed ET Symmetrical group Unsymmetrical group P
RB1
RB2
R3BB
Total
737 (64.0%)
558 (48.5%)
347 (30.1%)
1151
58 (35.4%)
38 (23.2%)
8 (4.9%)
164
<0.01
<0.01
<0.01
367 (58.5%)
248 (39.6%)
144 (23.0%)
627
22 (32.4%)
10 (14.7%)
3 (4.4%)
68
<0.01
<0.01
<0.01
Vol. 96., No. 3, Supplement, September 2011
CONCLUSION: Our results suggest that blastomere symmetry is an important predictor of blastocyst development and is sufficient for morphological selection of cleavage stage embryos.
P-488 Wednesday, October 19, 2011 RENEWING OF MEDIUM IN A SINGLE STEP MEDIA CULTURE PROTOCOL PROVIDES NO ADVANTAGE TO MOUSE EMBRYO DEVELOPMENT WHEN OBSERVED CONTINUOUSLY THROUGH TIME LAPSE MICROIMAGERY. M. D. VerMilyea, J. R. Graham, M. J. Tucker. Shady Grove Fertility Reproductive Science Center, Rockville, MD. OBJECTIVE: Extended embryo culture and single blastocyst transfer has resulted in the reformulation and development of new in-vitro culture media. Sequential media protocols require a disruption of embryo culture whereas medium in a non-interrupted single step medium (SSM) protocol is used continuously. There has been debate as to whether refreshing of SSM may be beneficial for embryo development; however, some argue against the disturbance to embryo(s) in culture. We applied time-lapse microimagery technology to the comparison of mouse embryos cultured in sequential, static and renewed culture media protocols. DESIGN: Prospective Animal Study. MATERIALS AND METHODS: Thawed 1-cell mouse embryos (Embryotech) were cultured individually in 25ml of either a prototype SSM (Irvine Scientific) or Quinn’s Advantage Cleavage and Blastocyst media (QA, Sage) at 37 C in reduced oxygen. Development was assessed using the EmbryoScope Monitoring System (Unisense, Fertilitech). Images were acquired at timed intervals for key morphologic stages as Mean Development Time (MDT). Traditional scoring of the degree of hatching (>50%) at 120hrs was also documented. RESULTS: The MDT of 1-cell embryos, cultured to the hatched blastocyst stage in assigned media were all similar (P>0.15, Student’s t-Test). At 120 hrs, by traditional scoring methods 69% (25/36) Sage, 61% (22/36) SSM and 55% (20/36) SSM Renewed were graded as >50% hatched (not significant, Sage vs. SSM (P¼0.62); Sage vs. SSM Renewed (P¼0.33); SSM vs. SSM Renewed (P¼0.81, Fisher’s exact). CONCLUSION: The detailed data on cleavage events and subsequent stages of development, provided by time-lapse imagery, show no statistical difference in MDT of 1-cell embryos cultured in Sage sequential, SSM or SSM renewed media. No differences in hatching rates, by traditional scoring, were also noted. Our results support the use of a SSM static protocol in IVF laboratories which may offer cost advantages and simplified culture management when compared to sequential culture media protocols.
P-489 Wednesday, October 19, 2011 EFFECTS OF ALPHA-LIPOIC ACID ON THE MOUSE EMBRYONIC DEVELOPMENT IN VITRO. J. Yoon, K. Juhn, S. Yoon, Y. Ko, J.-H. Lim. Maria Research Center, Seoul, Republic of Korea; Korea University, Seoul, Republic of Korea; Maria Fertility Hospital, Seoul, Republic of Korea. OBJECTIVE: Alpha-lipoic acid (LA) was reported to reduce the level of extracellular reactive oxygen species, which resulted in increase in the cell number of blastocyst stage embryos (Linck et al., 2007). The objective of the present study was to determine optimal concentration of LA when supplemented to culture medium under either low or high oxygen tension. DESIGN: Experimental animal study MATERIALS AND METHODS: Zygotes derived from BD F1 mice were developed in MRC#D01 medium (Biosupply Co., Korea) in the absence or presence of LA (0, 1, 10, 100 mM) under either 5% O2, 6% CO2 and 89% N2 or 6% CO2 in air. Embryo development was monitored on day 4 and 5. Efficacy of LA supplementation for blastocyst development was compared using a t-test and data were analyzed by SPSS. RESULTS: Regardless of O2 tension, 100 mM of LA significantly reduced the development of expanding blastocysts on day 4 and hatching or more developed blastocysts on day 5. Under 20% O2 tension, 1-10 mM of LA were significantly effective for the development of hatching or more developed blastocysts on day 5 (P<0.05), but not for the expanding or more developed ones on day 4. Under 5% O2 tension, however, 10 mM of LA significantly decreased the blastocyst development on both day 4 and 5 (P<0.05).
FERTILITY & STERILITYÒ
The effect of LA concentrations on the mouse embryo development in vitro
Oxygen tension
LA concentration (mM)
No. of zygotes examined
R expanding blastocyst (day 4)
R hatching blastocyst (day 5)
20% O2 20% O2 20% O2 20% O2 5% O2 5% O2 5% O2 5% O2
0 1 10 100 0 1 10 100
228 225 218 220 144 144 136 140
90.5 3.9a 94.0 4.2a 92.9 2.3a 80.9 1.3b 97.0 4.7c 94.7 4.1c 88.3 3.9d 78.1 9.6e
72.6 16.2f 92.8 3.7g 91.8 5.7g 70.8 3.0f 94.4 4.4h 94.2 4.5h 88.3 3.9i 75.5 12.7i
a-b, c-d, c-e, d-e, f-g, h-i indicate significant differences in the column (P<0.05 in t-test) CONCLUSION: Data from the present study indicate that 1 mM of LA supplementation to culture medium is practically useful for mouse embryonic development under both low and high oxygen tensions.
OVARIAN STIMULATION P-490 Wednesday, October 19, 2011 SUCCESS OF CLOMIPHENE CITRATE IN A COUNTY POPULATION. E. Mellano, K. Brennan, C. Holschneider. Obstetrics and Gynecology, University of California Los Angeles Medical Center, Los Angeles, CA; Obstetrics and Gynecology, Olive View-UCLA Medical Center, Sylmar, CA. OBJECTIVE: To determine the effectiveness of clomiphene citrate (CC) and timed intercourse (TI) in achieving pregnancy in an underserved county population. DESIGN: Retrospective analysis. MATERIALS AND METHODS: The medical records of all women who presented to a metropolitan public hospital for infertility between September 2010 and January 2011 and were prescribed CC for ovulation induction or superovulation were analyzed. Data was collected on the 35 women identified including age, body mass index (BMI), infertility diagnosis, day 3 follicle stimulating hormone (FSH) and estradiol (E2) levels, semen analysis, hysterosalpingogram (HSG) results, number of CC cycles, day 21 and 28 progesterone (P4) and BhCG, with pregnancy defined as a positive BhCG. RESULTS: Of the 35 patients, 31 had normal semen analyses and HSGs, and known ovulatory status and BhCG results of at least one cycle. The total number of CC cycles was 86; of these, 58 were ovulatory as defined by P4 > 3 ng/ml and 2 pregnancies were identified. The ovulation rate per CC cycle was 67.4%. The pregnancy rate per ovulatory cycle was 3.45%. The patients were then subdivided by those who always ovulated (16/31); those who sometimes ovulated (6/31); and those who never ovulated (9/31); no significant differences regarding age, day 3 E2 or FSH were found. However, women with a higher BMI and with polycystic ovarian syndrome or oligomenorrhea were less likely to ovulate with CC (P<0.005). CONCLUSION: The majority of patients ovulated with CC, but few became pregnant. Of the two documented pregnancies, one was an ectopic and the other was lost to follow-up. In this county population, CC and TI alone are not an effective treatment for infertility. Adjuncts, such as dexamethasone or metformin, may increase success in this underserved population. Supported by: Departmental
P-491 Wednesday, October 19, 2011 DOES EARLIER ADMINISTRATION OF HUMAN CHORIONIC GONADOTROPIN (hCG) IMPROVE THE PROBABILITY OF PREGNANCY IN CYCLES STIMULATED WITH REC-FSH AND GnRH ANTAGONISTS? A PROSPECTIVE RANDOMIZED TRIAL. D. Kyrou, E. M. Kolibianakis, H. M. Fatemi, B. C. Tarlatzis, H. Tournaye, P. Devroey. Centre for Reproductive Medicine, Universitair Ziekenhuis Brussel, Brussels, Belgium. OBJECTIVE: The purpose of this trial was to assess the effect of altering the timing of hCG administration on the probability of pregnancy in patients stimulated with rec-FSH/GnRH antagonists for IVF.
S251
P-485 Wednesday, October 19, 2011 EFFECT OF EMBRYO GROUP CULTURE STRATEGY ON THE BLASTOCYST DEVELOPMENT AND PREGNANCY OUTCOME. T. Tao, A. Robichaud, J. Mercier, R. Ouellette. Conceptia Clinic, Moncton, NB, Canada. OBJECTIVE: The aim of this study is to investigate the influence of different embryo group culture strategy on the blastocyst development and pregnancy outcome. DESIGN: Two embryo group culture methods were compared. MATERIALS AND METHODS: Oocytes were inseminated either by standard IVF or ICSI. All zygotes were individually cultured in 20 ml droplets of G1.3 medium under oil at 37 C, in a humidified atmosphere containing 6% CO2, 5% O2 and 89 % N2 until day 3. On this day embryos were classified into two categories: good quality (have even, regular and spherical blastomeres, with R6 cells and %15% fragmentation) and poor quality. Embryos were then cultured in group of 2-5 embryos per droplet (50 ml G2.3) until day 5 or 6. The group culture strategy is: A, embryos were randomly grouped regardless of embryo quality; B, good or poor quality of embryos was separately grouped. Maximum of two blastocysts were transferred to patient on day 5. RESULTS: Results were shown in table 1. The blastocyst rate (62%) in group B were significantly higher than that in group A (43%). There was no statistical difference between two groups in the pregnancy rate. TABLE 1. Outcome of blastocyst development and pregnancy
Group # Patient A B
36 31
# embryos Average embryos cultured # Blastocysts transferred # Pregnancy 397 291
172 (43%) a 179 (62%) b
1.8 1.9
26 (72%) 20 (65%)
In group B, the blastocyst rates for good or poor quality of day 3 embryos were, respectively, 74% (159/215) and 26% (20/76), the average was 62%. a,bValues with different superscript letters within the same column are significant different (P<0.05). CONCLUSION: Our results suggest that day 3 embryos with good or poor quality should be separately grouped for culture to blastocyst stage. Autocrine or paracrine growth or survival factors produced by the embryos may support itself and other embryos in their development. Some other unknown factors released from poor quality embryos (such as arrested embryos) may inhibit the surrounding embryo development.
P-486 Wednesday, October 19, 2011 METABOLOMIC PROFILE OF EMBRYO CULTURE MEDIA UNDER HYPOXIA. J. M. De los Santos, P. Gamiz, F. Domınguez, A. Galan, S. Perez, M. J. De los Santos. IVF Laboratory, IVI Valencia, INCLIVA, Valencia, Spain; Embryomics S.L., Derio, Vizcaya, Spain. OBJECTIVE: Aim of this study was to determine whether human pre-embryos cultured under different oxygen concentrations (5% vs 20%) suffer from some sort of change in their metabolism.
S250
Abstracts
DESIGN: Prospective study. MATERIALS AND METHODS: A total of 23 patients of our ovum donation program were incluided in this study. 41 samples of spent embryo culture media (n ¼ 24; 5% [O2]; n ¼ 17; 20% [O2]) were used. Both groups selected had 100% of implantation rate. Embryos were cultured in O2/CO2 incubator. Culture media and controls were collected on day +3 and stored at -186 C. There were no differences on the age of the donors (26.2 vs 25.8), neither on the embryo quality. Samples were thawed and metabolites were extracted, then injected into ultra performance liquid chromatograph coupled to a mass spectrometer. All endogenous metabolites present in samples were measured, defined by its relation mass/charge and their retention time. For statistical analysis, univariate analysis (comparison of means) of each metabolite, between both groups and multivariate (principal component analysis) to find overall differences between groups were made. P values <0.05 were considered significant. RESULTS: Multivariate analysis of the global metabolomic profile of embryos cultured in hypoxia vs atmospheric conditions did not reveal significant differences between both groups. Despite no differences were observed in global profile, through univariate analysis, two metabolites defined by their relationship mass/charge and retention time, appeared significantly ‘‘high’’, while a third was diminished, in the group of embryos grown under low oxygen tensions. CONCLUSION: Culture of human pre-embryos in low oxygen tension did not change significantly their metabolic glycolysis pathways during in vitro development because there were no changes in the consumption profile of glucose, pyruvate or lactate. Furthermore, amino acids used as implantation markers such as Asn, Gly or Leu are not altered under both oxygen culture conditions.
P-487 Wednesday, October 19, 2011 BLASTOMERE SYMMETRY IS AN IMPORTANT PREDICTOR OF BLASTOCYST DEVELOPMENT. K. Nakayama, N. Fukunaga, R. Nagai, H. Kitasaka, Y. Hashiba, Y. Asada. Asada Ladies Clinic, Nagoya, Aichi, Japan; The Asada Institute for Reproductive Medicine, Kasugai, Aichi, Japan. OBJECTIVE: The aim of this study was to evaluate whether the blastomere symmetry of day 3 embryos affects blastocyst development on day 3. DESIGN: Retrospective study. MATERIALS AND METHODS: We reviewed the outcome of Day 5 embryo transfer (ET) using fresh embryos (436 cycles) and frozen and thawed embryos (489 cycles) between January 1, 2008 and December 31, 2009. Blastocyst development was assessed for embryos divided into two groups based on morphology: symmetrical group (uniform blastomeres) and unsymmetrical group (uneven blastomeres). All embryos had 6-8 cells and %20% fragmentation on Day 3. Chi-square test was used to examine the differences between the groups. RESULTS: Blastosyst development was significantly higher for the symmetrical group than the unsymmetrical group at all score levels for ET performed with fresh and frozen and thawed embryos (Table 1).
TABLE 1. Results of Day 5 ET for fresh and frozen and thawed embryos
Gardner’s score
fresh ET Symmetrical group Unsymmetrical group P frozen-thawed ET Symmetrical group Unsymmetrical group P
RB1
RB2
R3BB
Total
737 (64.0%)
558 (48.5%)
347 (30.1%)
1151
58 (35.4%)
38 (23.2%)
8 (4.9%)
164
<0.01
<0.01
<0.01
367 (58.5%)
248 (39.6%)
144 (23.0%)
627
22 (32.4%)
10 (14.7%)
3 (4.4%)
68
<0.01
<0.01
<0.01
Vol. 96., No. 3, Supplement, September 2011
CONCLUSION: Our results suggest that blastomere symmetry is an important predictor of blastocyst development and is sufficient for morphological selection of cleavage stage embryos.
P-488 Wednesday, October 19, 2011 RENEWING OF MEDIUM IN A SINGLE STEP MEDIA CULTURE PROTOCOL PROVIDES NO ADVANTAGE TO MOUSE EMBRYO DEVELOPMENT WHEN OBSERVED CONTINUOUSLY THROUGH TIME LAPSE MICROIMAGERY. M. D. VerMilyea, J. R. Graham, M. J. Tucker. Shady Grove Fertility Reproductive Science Center, Rockville, MD. OBJECTIVE: Extended embryo culture and single blastocyst transfer has resulted in the reformulation and development of new in-vitro culture media. Sequential media protocols require a disruption of embryo culture whereas medium in a non-interrupted single step medium (SSM) protocol is used continuously. There has been debate as to whether refreshing of SSM may be beneficial for embryo development; however, some argue against the disturbance to embryo(s) in culture. We applied time-lapse microimagery technology to the comparison of mouse embryos cultured in sequential, static and renewed culture media protocols. DESIGN: Prospective Animal Study. MATERIALS AND METHODS: Thawed 1-cell mouse embryos (Embryotech) were cultured individually in 25ml of either a prototype SSM (Irvine Scientific) or Quinn’s Advantage Cleavage and Blastocyst media (QA, Sage) at 37 C in reduced oxygen. Development was assessed using the EmbryoScope Monitoring System (Unisense, Fertilitech). Images were acquired at timed intervals for key morphologic stages as Mean Development Time (MDT). Traditional scoring of the degree of hatching (>50%) at 120hrs was also documented. RESULTS: The MDT of 1-cell embryos, cultured to the hatched blastocyst stage in assigned media were all similar (P>0.15, Student’s t-Test). At 120 hrs, by traditional scoring methods 69% (25/36) Sage, 61% (22/36) SSM and 55% (20/36) SSM Renewed were graded as >50% hatched (not significant, Sage vs. SSM (P¼0.62); Sage vs. SSM Renewed (P¼0.33); SSM vs. SSM Renewed (P¼0.81, Fisher’s exact). CONCLUSION: The detailed data on cleavage events and subsequent stages of development, provided by time-lapse imagery, show no statistical difference in MDT of 1-cell embryos cultured in Sage sequential, SSM or SSM renewed media. No differences in hatching rates, by traditional scoring, were also noted. Our results support the use of a SSM static protocol in IVF laboratories which may offer cost advantages and simplified culture management when compared to sequential culture media protocols.
P-489 Wednesday, October 19, 2011 EFFECTS OF ALPHA-LIPOIC ACID ON THE MOUSE EMBRYONIC DEVELOPMENT IN VITRO. J. Yoon, K. Juhn, S. Yoon, Y. Ko, J.-H. Lim. Maria Research Center, Seoul, Republic of Korea; Korea University, Seoul, Republic of Korea; Maria Fertility Hospital, Seoul, Republic of Korea. OBJECTIVE: Alpha-lipoic acid (LA) was reported to reduce the level of extracellular reactive oxygen species, which resulted in increase in the cell number of blastocyst stage embryos (Linck et al., 2007). The objective of the present study was to determine optimal concentration of LA when supplemented to culture medium under either low or high oxygen tension. DESIGN: Experimental animal study MATERIALS AND METHODS: Zygotes derived from BD F1 mice were developed in MRC#D01 medium (Biosupply Co., Korea) in the absence or presence of LA (0, 1, 10, 100 mM) under either 5% O2, 6% CO2 and 89% N2 or 6% CO2 in air. Embryo development was monitored on day 4 and 5. Efficacy of LA supplementation for blastocyst development was compared using a t-test and data were analyzed by SPSS. RESULTS: Regardless of O2 tension, 100 mM of LA significantly reduced the development of expanding blastocysts on day 4 and hatching or more developed blastocysts on day 5. Under 20% O2 tension, 1-10 mM of LA were significantly effective for the development of hatching or more developed blastocysts on day 5 (P<0.05), but not for the expanding or more developed ones on day 4. Under 5% O2 tension, however, 10 mM of LA significantly decreased the blastocyst development on both day 4 and 5 (P<0.05).
FERTILITY & STERILITYÒ
The effect of LA concentrations on the mouse embryo development in vitro
Oxygen tension
LA concentration (mM)
No. of zygotes examined
R expanding blastocyst (day 4)
R hatching blastocyst (day 5)
20% O2 20% O2 20% O2 20% O2 5% O2 5% O2 5% O2 5% O2
0 1 10 100 0 1 10 100
228 225 218 220 144 144 136 140
90.5 3.9a 94.0 4.2a 92.9 2.3a 80.9 1.3b 97.0 4.7c 94.7 4.1c 88.3 3.9d 78.1 9.6e
72.6 16.2f 92.8 3.7g 91.8 5.7g 70.8 3.0f 94.4 4.4h 94.2 4.5h 88.3 3.9i 75.5 12.7i
a-b, c-d, c-e, d-e, f-g, h-i indicate significant differences in the column (P<0.05 in t-test) CONCLUSION: Data from the present study indicate that 1 mM of LA supplementation to culture medium is practically useful for mouse embryonic development under both low and high oxygen tensions.
OVARIAN STIMULATION P-490 Wednesday, October 19, 2011 SUCCESS OF CLOMIPHENE CITRATE IN A COUNTY POPULATION. E. Mellano, K. Brennan, C. Holschneider. Obstetrics and Gynecology, University of California Los Angeles Medical Center, Los Angeles, CA; Obstetrics and Gynecology, Olive View-UCLA Medical Center, Sylmar, CA. OBJECTIVE: To determine the effectiveness of clomiphene citrate (CC) and timed intercourse (TI) in achieving pregnancy in an underserved county population. DESIGN: Retrospective analysis. MATERIALS AND METHODS: The medical records of all women who presented to a metropolitan public hospital for infertility between September 2010 and January 2011 and were prescribed CC for ovulation induction or superovulation were analyzed. Data was collected on the 35 women identified including age, body mass index (BMI), infertility diagnosis, day 3 follicle stimulating hormone (FSH) and estradiol (E2) levels, semen analysis, hysterosalpingogram (HSG) results, number of CC cycles, day 21 and 28 progesterone (P4) and BhCG, with pregnancy defined as a positive BhCG. RESULTS: Of the 35 patients, 31 had normal semen analyses and HSGs, and known ovulatory status and BhCG results of at least one cycle. The total number of CC cycles was 86; of these, 58 were ovulatory as defined by P4 > 3 ng/ml and 2 pregnancies were identified. The ovulation rate per CC cycle was 67.4%. The pregnancy rate per ovulatory cycle was 3.45%. The patients were then subdivided by those who always ovulated (16/31); those who sometimes ovulated (6/31); and those who never ovulated (9/31); no significant differences regarding age, day 3 E2 or FSH were found. However, women with a higher BMI and with polycystic ovarian syndrome or oligomenorrhea were less likely to ovulate with CC (P<0.005). CONCLUSION: The majority of patients ovulated with CC, but few became pregnant. Of the two documented pregnancies, one was an ectopic and the other was lost to follow-up. In this county population, CC and TI alone are not an effective treatment for infertility. Adjuncts, such as dexamethasone or metformin, may increase success in this underserved population. Supported by: Departmental
P-491 Wednesday, October 19, 2011 DOES EARLIER ADMINISTRATION OF HUMAN CHORIONIC GONADOTROPIN (hCG) IMPROVE THE PROBABILITY OF PREGNANCY IN CYCLES STIMULATED WITH REC-FSH AND GnRH ANTAGONISTS? A PROSPECTIVE RANDOMIZED TRIAL. D. Kyrou, E. M. Kolibianakis, H. M. Fatemi, B. C. Tarlatzis, H. Tournaye, P. Devroey. Centre for Reproductive Medicine, Universitair Ziekenhuis Brussel, Brussels, Belgium. OBJECTIVE: The purpose of this trial was to assess the effect of altering the timing of hCG administration on the probability of pregnancy in patients stimulated with rec-FSH/GnRH antagonists for IVF.
S251