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ICSI therapy
P-361 THE EFFECT OF COTININE ON SPERMATOZOA QUALITY AND OUTCOME OF PATIENTS UNDERGOING IVF/ICSI THERAPY. M. E. Hammadeh, C. Fischer-Hammadeh, J. Al-Qatami, M. F. Hamad, P. Rosenbaum, W. Schmidt. Obstetrics and Gynecology, University of Saarland, Homburg/Saar, Germany; University of Saarland, Homburg/Saar, Germany. OBJECTIVE: The objectives of this study were to determine the relationship between reactive oxygen species (ROS), total antioxidant (TAS) and Cotinine levels in seminal plasma and sperm parameters as well as their effect on IVF/ICSI outcome. DESIGN: Prospective study. MATERIALS AND METHODS: Semen concentration of ROS, TAS and sperm parameters concentration, motility, motility, vitality, membrane integrity (HOS-test), morphology, chromatin integrity (CMA3), DNA Fragmentation (Tunel-test) of 46 patients (11) heavy smokers and (35) non smokers were investigated. As semen liquefication and assessment, the semen was separated from seminal plasma using PureSperm gradient centrifugation. The seminal plasma on the top of the upper layer of the two fraction 45% and 90% of PureSperm layer was withdrawn after centrifugation for sperm preparation and kept frozen at - 80 C until the concentrations of ROS, TAS, and Cotinine were measured by colorimetric assay using commercially available kits. Cotinine concentration was measured with (ELISA) kit. The motile sperm which obtained after sperm preparation was used for injection (ICSI) or insemination of Oocytes (IVF). RESULTS: The sperm concentration, mean number of motile spermatozoa after sperm preparation, morphologically normal spermatozoa, and mean number of cleaved and transferred Oocytes and pregnancy rate were significantly higher in non-smoking patients group in comparison to smoking one (76.229.5mill/ml; 65.231.4%; 16.118.5%; 1.90.8%; 34%vs.50.739.2 mill/ml; 39.020.4%19.818.5%; 1.90.7% and 27.3% respectively). The other sperm parameters were similar in both groups. Cotinine concentration was significantly higher in smokers than non-smokers (144.382.2mg/dl vs.19.323.7mg/dl; p¼0.001). Inverse correlations were found between Cotinine concentration and ejaculate volume, sperm concentration, motility, vitality, morphology membrane integrity, DNA fragmentation (TUNEL), TAS concentration and fertilization. Besides, a positive correlation was shown between Cotinine and ROS concentration. In the smoker group three patients become pregnant (27.3%) and 11 patients achieved pregnancy in the non-smoking group (34%; p¼0.001). CONCLUSIONS: Cotinine concentration in seminal plasma has an adverse effect on seminal plasma anti oxidants, sperm parameters and ART outcome and increase the production of ROS in seminal plasma. Therefore, it is recommended to quit smoking in childless patients who are seeking ART therapy. Supported by: University of Saarland budget only.
P-362 OOCYTE MATURITY: DOES IT PREDICT IVF OUTCOME? A REVIEW OF A LARGE IVF PROGRAM. J. Liebermann, A. N. Beltsos, E. J. Pelts, J. M. Matthews, M. Uhler, D. D. Ezcurra. Fertility Centers of Illinois, Chicago, IL; Fertility Centers of Illinois, Chicago; Medical Affairs and Reproductive Health, EMD Serono Inc., Rockland, MA. OBJECTIVE: In general, embryologists consider that a reduction in the oocyte maturity (mature oocytes/total oocytes retrieved) reduces the chances of producing more total embryos (freshþfrozen) as well as reduces the chance of achieving pregnancy. Therefore, to prove this hypothesis, two variables were evaluated: 1) total number of oocytes retrieved and 2) percentage of those that are at the mature stage (Metaphase II) with an extruded first polar body and a good cytoplasmatic morphology. DESIGN: Retrospective observational study. MATERIALS AND METHODS: 1538 retrievals from the entire population of patients undergoing IVF from January 1st to December 31st, 2007, were included in this analysis. Retrievals were stratified according to the total number of oocytes retrieved into three groups: 1-10, 10-20 and 20-30 as well as according to the rate of maturity: 60-70%, 70-80% and 80-100%, considering that, in our hands, these were the most frequent percentages of MII’s seen. Standard laboratory protocols were followed, including intracytoplas-
S228
Abstracts
mic sperm injection, assisted hatching for day 3 transfers and extended culture for blastocyst transfers. Statistical analysis was done using ANOVA test for continuous data and Chi Square test for categorical data; statistical significance was set at p<0.05. RESULTS: The characteristics and outcomes of 1538 retrievals were presented (Table 1). The results expressed as mean SD or percentages, as appropriate, are summarized below. When the number of oocytes retrieved increased, there was a decrease in the number of MII but an increase in blastocyst development as well as clinical pregnancy rates. Maturity rate does not appear to be predictive of outcome. TABLE 1. Total
Percentage of
Oocytes Retrieved
MII (%)
1-10
11-20
21-30
60-70
70-80
80-100
N
512
695
331
257
290
# of MII (%)
77*
75
75
-
-
# of oocytes retrieved Fertilization rate (%)
-
-
-
14.48.7
15.68.4
672 13.2±7.9*
68*
70
73
68
69
Cleavage rate (%)
95
97
96
95
99
96
# of Day 3 embryos
65
68
68
64
70
68
72*
with R6 cells (%) Blastocyst rate (%) Total embryos produced
19*
47
57
41
43
44
2.0±1.8*
5.64.5
128.7
5.87.1
6.45.8
6.66.6
28%*
47%
54%
43%
46%
44%
(freshþfrozen) Clinical pregnancy (%)
(144)
(328)
(179)
(110)
(135)
(300)
* Variables with P-value <0.05, all the other NS. CONCLUSIONS: When considering the predictive value of MII ratio to predict successful outcome of Controlled Ovarian Stimulation, the total amount of oocytes retrieved is important. Making over 10 oocytes improves chance of positive outcome. Supported by: None.
P-363 MORE THAN 9 MM OF ENDOMETRIUM THICKNESS AT THE DAY OF HCG TRIGGERING PREDICTS BETTER IVF OUTCOMES OF THE PATIENTS WITH DAY 5 EMBRYO TRANSFER. S. Soliman, R. Baydoun, L. Al-Qahwaji, J. Xu, B. Wang. Newlife Fertility Center, Mississauga, ON, Canada. OBJECTIVE: The clinical IVF data is examined to correlate endometrium thickness at the day of HCG triggering with the outcomes of IVF cycles in the patients with day 5 blastocyst transfers. DESIGN: Retrospective clinical data analysis. MATERIALS AND METHODS: The IVF data of the patients (age from 24 to 42.2) enrolled in day 5 blastocyst transfer program in 2007 is pooled and subjected to statistical analysis with Graphpad Prism 5. The patients were given various hormonal stimulation regimes. The endometrium thickness was measured at the day of HCG injection. Retrievals were conducted 38 hours after HCG injection. Recovered eggs were fertilized with ICSI or insemination depending on the parameters of sperm analysis. The embryos were cultured with the sequential media in 5% O2, 6% CO2 at 37 oC for 5 days. Good quality of blastocysts were selected for intrauterine transfer under the ultrasonic guidance. The pregnancies were detected by the levels of HCG at the day 11-12 following embryo transfers and further confirmed weeks later by fetal heart beats. The patients were grouped according to the thickness of endometrium, various endpoints (listed in the table) were compared among the different groups. The difference of percentage data was examined by Fisher’s exact test and means were analysed by students T test. RESULTS: See results in table 1. CONCLUSIONS: The patients with above 9 mm of endometrium at the day of HCG triggering resulted in better IVF outcomes with day 5 blastocyst transfers. Endometrium thickness is a critical parameter for predicting the outcomes of an IVF cycle of day 5 blastocyst transfer.
Vol. 90, Suppl 1, September 2008
P-362 OOCYTE MATURITY: DOES IT PREDICT IVF OUTCOME? A REVIEW OF A LARGE IVF PROGRAM. J. Liebermann, A. N. Beltsos, E. J. Pelts, J. M. Matthews, M. Uhler, D. D. Ezcurra. Fertility Centers of Illinois, Chicago, IL; Fertility Centers of Illinois, Chicago; Medical Affairs and Reproductive Health, EMD Serono Inc., Rockland, MA. OBJECTIVE: In general, embryologists consider that a reduction in the oocyte maturity (mature oocytes/total oocytes retrieved) reduces the chances of producing more total embryos (freshþfrozen) as well as reduces the chance of achieving pregnancy. Therefore, to prove this hypothesis, two variables were evaluated: 1) total number of oocytes retrieved and 2) percentage of those that are at the mature stage (Metaphase II) with an extruded first polar body and a good cytoplasmatic morphology. DESIGN: Retrospective observational study. MATERIALS AND METHODS: 1538 retrievals from the entire population of patients undergoing IVF from January 1st to December 31st, 2007, were included in this analysis. Retrievals were stratified according to the total number of oocytes retrieved into three groups: 1-10, 10-20 and 20-30 as well as according to the rate of maturity: 60-70%, 70-80% and 80-100%, considering that, in our hands, these were the most frequent percentages of MII’s seen. Standard laboratory protocols were followed, including intracytoplas-
S228
Abstracts
mic sperm injection, assisted hatching for day 3 transfers and extended culture for blastocyst transfers. Statistical analysis was done using ANOVA test for continuous data and Chi Square test for categorical data; statistical significance was set at p<0.05. RESULTS: The characteristics and outcomes of 1538 retrievals were presented (Table 1). The results expressed as mean SD or percentages, as appropriate, are summarized below. When the number of oocytes retrieved increased, there was a decrease in the number of MII but an increase in blastocyst development as well as clinical pregnancy rates. Maturity rate does not appear to be predictive of outcome. TABLE 1. Total
Percentage of
Oocytes Retrieved
MII (%)
1-10
11-20
21-30
60-70
70-80
80-100
N
512
695
331
257
290
# of MII (%)
77*
75
75
-
-
# of oocytes retrieved Fertilization rate (%)
-
-
-
14.48.7
15.68.4
672 13.2±7.9*
68*
70
73
68
69
Cleavage rate (%)
95
97
96
95
99
96
# of Day 3 embryos
65
68
68
64
70
68
72*
with R6 cells (%) Blastocyst rate (%) Total embryos produced
19*
47
57
41
43
44
2.0±1.8*
5.64.5
128.7
5.87.1
6.45.8
6.66.6
28%*
47%
54%
43%
46%
44%
(freshþfrozen) Clinical pregnancy (%)
(144)
(328)
(179)
(110)
(135)
(300)
* Variables with P-value <0.05, all the other NS. CONCLUSIONS: When considering the predictive value of MII ratio to predict successful outcome of Controlled Ovarian Stimulation, the total amount of oocytes retrieved is important. Making over 10 oocytes improves chance of positive outcome. Supported by: None.
P-363 MORE THAN 9 MM OF ENDOMETRIUM THICKNESS AT THE DAY OF HCG TRIGGERING PREDICTS BETTER IVF OUTCOMES OF THE PATIENTS WITH DAY 5 EMBRYO TRANSFER. S. Soliman, R. Baydoun, L. Al-Qahwaji, J. Xu, B. Wang. Newlife Fertility Center, Mississauga, ON, Canada. OBJECTIVE: The clinical IVF data is examined to correlate endometrium thickness at the day of HCG triggering with the outcomes of IVF cycles in the patients with day 5 blastocyst transfers. DESIGN: Retrospective clinical data analysis. MATERIALS AND METHODS: The IVF data of the patients (age from 24 to 42.2) enrolled in day 5 blastocyst transfer program in 2007 is pooled and subjected to statistical analysis with Graphpad Prism 5. The patients were given various hormonal stimulation regimes. The endometrium thickness was measured at the day of HCG injection. Retrievals were conducted 38 hours after HCG injection. Recovered eggs were fertilized with ICSI or insemination depending on the parameters of sperm analysis. The embryos were cultured with the sequential media in 5% O2, 6% CO2 at 37 oC for 5 days. Good quality of blastocysts were selected for intrauterine transfer under the ultrasonic guidance. The pregnancies were detected by the levels of HCG at the day 11-12 following embryo transfers and further confirmed weeks later by fetal heart beats. The patients were grouped according to the thickness of endometrium, various endpoints (listed in the table) were compared among the different groups. The difference of percentage data was examined by Fisher’s exact test and means were analysed by students T test. RESULTS: See results in table 1. CONCLUSIONS: The patients with above 9 mm of endometrium at the day of HCG triggering resulted in better IVF outcomes with day 5 blastocyst transfers. Endometrium thickness is a critical parameter for predicting the outcomes of an IVF cycle of day 5 blastocyst transfer.
Vol. 90, Suppl 1, September 2008