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β2 receptor agonists during the peri-implantation period could improve implantation and pregnancy rates.
a deletion of the short arm of one of the X-chromosomes (46 X, del (X) (p22.1). Subsequently, a decision was made to proceed with donor oocyte IVF and blastocyst transfer. The oocyte donor was stimulated with recombinant FSH for 8 days, coasted for three days and hCG administered on day 11 of stimulation. 22 mature oocytes were retrieved and then sperm injected due to low sperm counts. Of the 20 fertilized oocytes, 2 quality expanded blastocysts were available for transfer on day 5. One of these blastocysts had two distinct ICM’s (Figure 1) and monozygotic twinning was predicted. Since no additional embryos were available for selection, both these embryos were transferred 118 h after ICSI. Results: A trichorionic heterozygotic triplet pregnancy with dichorionic, monozygotic twins was diagnosed 29 days post oocyte retrieval (Figure 2). At this time, three heartbeats were noted. Triplet B was reduced 3 months post oocyte retrieval. Triplet C died in utero approximately 4 months after the oocyte retrieval. Triplet A is ongoing at 33 weeks at the time of writing.
- 300 IU/day of r-FSH were administered subcutaneously for the first 7 days for ovarian stimulation and ultrasound control was started on the 8th day, with adaptation of the dose according to the ovarian response. Results: Clinical and laboratory details of the studied groups are shown in Table 1.
No. of transfers Age (years) Oocytes collected MII fertilization rate Embryo escore Implantation rate (%) Pregnancy per transfer (%) Abortion (%)
Control
Terbutaline
Ritodrine
P
25 34.8 ⫾ 0.87 10.2 ⫾ 1.4 66.9 ⫾ 4.7 29.1 ⫾ 2.8 15.7 (11/70) 32 (8/25) 12.5
47 33.6 ⫾ 0.66 12.1 ⫾ 1.1 65.5 ⫾ 3.3 28.7 ⫾ 1.8 17.2 (22/128) 34 (16/47) 25
43 34.5 ⫾ 0.73 11.4 ⫾ 1.0 75 ⫾ 3.1 30.8 ⫾ 2.1 15 (18/120) 30.2 (13/43) 15.4
NS NS NS NS NS NS NS
Conclusions: Implantation and pregnancy rates are slightly higher in the terbutaline group (17.2%, 34%) than in ritodrine group (15%, 30.2%) and control (15.7%, 32%). However, at the present time this differences are not statistically significant.
P-183 Telomerase activity in human blastomeres: a predictor for embryo viability and development? C. Wang, S. Horng, C. Chang, H. Huang, H. Wang, Y. Soong. Chang Gung Memorial Hosp, Kweishan, Taoyuan, Taiwan.
Conclusions: In this program, monochorionic, diamnionic placentation is the most common placentation type observed in monozygotic pregnancies resulting from blastocyst transfers (19/27 patients). According to literature, a double ICM blastocyst should have resulted in a similar monochorionic twin. The occurrance of a dichorionic twin may imply that, in this case, the trophoblast cells were already destined to separate at the time of observation or that the separation could not be observed. Since all of this occurred well before embryo hatching, this case of monozygotic twinning can not be explained by in vitro zona alteration. The occurrance of two ICM’s in the same blastocyst demonstrates at least one likely mechanism by which monozygotic twinning may result when transferring blastocysts. P-182
2 receptor agonists during the peri-implantation period could improve implantation and pregnancy rates. J. G. Franco Jr, R. L. Baruffi, E. Garbellini, V. Felipe, C. Petersen, A. L. Mauri. Ctr for Human Reproduction Sinha´ Junqueira Maternity Fdn, Ribeirao Preto, Brazil. Objective: 2 receptor agonists can decrease uterine activity and increased endometrial blood flow (Akerlund and Andersson, 1976). On the other hand, decreased uterine contractions frequency at the time of embryo transfer is associated with increased implantation and pregnancy rates (Fanchin et al., 1998). Impaired perfusion of endometrial blood flow has an unfavourable reproductive outcome (Yang et al., 1999). The purpose of this study was to compare in a prospective randomized trial the use of two b2 receptor agonists (terbutaline and ritodrine) during the peri-implantation period in terms of the ICSI program outcome. Design: A prospective randomised clinical study. Materials/Methods: A total of 115 patients from ICSI program were random assigned to one of the drugs (2/1) by drawing lots, using a randomization table previously elaborated for the study. The terbutaline group received 10 mg/day for 15 days from the day after oocyte collection up to the pregnancy test (14 days post-embryo transfer). The ritodrine group received 20 mg/day for the same period of time terbutaline group. All women were stimulated for oocyte retrieval with r-FSH (Puregon®) after blockade of as the 2nd phase (starting on the 21st day of the cycle) with nasal nafarelin (Synarel®) at the dose of 400 g a day until the day of administration of human chorionic gonadotropin (hCG). Fixed doses of 150
FERTILITY & STERILITY威
Objective: Telomerase is a ribonucleoprotein that synthesizes telomeric DNA at the ends of eukaryotic chromosomes. It has been hypothesized that telomerase activity is necessary for cellular immortalization and that telomerase activity is present in cells of germline origin and embryos. Consistent with the hypothesis, telomerase activity has been detected in extracts of human ovaries and testes. But it has not been measured in single human blastomere because of difficulty in obtaining sufficient DNA for analysis. Based on the importance of telomerase for meiosis and mitosis, we designed this study for the assessment of telomerase activity in single blastomeres with the attempt to establish a possible selection or scoring system, in order to select a better embryo for uterine transfer, or to be used as a marker for embryonic health. Design: Prospective clinical study with RT-PCR for telomerase activity in individual human blastomeres. Materials/Methods: Oocytes were obtained from patients undergoing an assisted fertilization procedure after ovarian stimulation with GnRH agonist in combination with FSH/ or HMG and hCG. IVF or ICSI were performed as routine. The fertilized oocytes were cultured for three days. Group A consisted of 21 normal cleaving 3PN embryos at 4 to 8 cell stage. Group B consisted of 35 arrested or fragmented embryos at 2 to 4 cell stage, which were considered to be compromised. After removing the zona pellucida. the embryos were placed in calcium- and magnesium-free medium for dissociation of blastomeres. RNA extraction, reverse transcription, and hTCS (human telomerase catalytic subunit) PCR amplication were performed. The amplification resulted in a PCR products size of 145 bp, then were separated by 3% gel electrophoresis. Siha cells and purified water were used as positive and negative control. Results: 95 intact blastomeres were dissociated from 21 embryos in Group A and 110 intact blastomeres from 35 embryos in Group B. Of these, 86 (90.5%) out of 95 blastomeres in Group A and 78 (70.9%) out of 110 blastomeres in Group B demonstrated telomerase activity by the expression of 145 bp product from hTCS mRNA. The difference was significant in statistics (P ⬍ 0.05, Chi-square). Conclusions: The limitations of evaluating embryos based on morphological criteria alone, as are usually done in most infertility centers, are well recognized. Correlations between gross morphology and implantation are weak and inaccurate. Although the results are not satisfactory, many studies have researched more objective criteria for judging embryo viability and implantation potential, in order to select better embryos to transfer, and consequently enhance the implantation and pregnancy rate without increasing the multiple pregnancies. To our knowledge, there is still no report regarding the relationship between telomerase activity and embryo quality. Our study was conducted to assess telomerase activity in individual blas-
S173
- 300 IU/day of r-FSH were administered subcutaneously for the first 7 days for ovarian stimulation and ultrasound control was started on the 8th day, with adaptation of the dose according to the ovarian response. Results: Clinical and laboratory details of the studied groups are shown in Table 1.
No. of transfers Age (years) Oocytes collected MII fertilization rate Embryo escore Implantation rate (%) Pregnancy per transfer (%) Abortion (%)
Control
Terbutaline
Ritodrine
P
25 34.8 ⫾ 0.87 10.2 ⫾ 1.4 66.9 ⫾ 4.7 29.1 ⫾ 2.8 15.7 (11/70) 32 (8/25) 12.5
47 33.6 ⫾ 0.66 12.1 ⫾ 1.1 65.5 ⫾ 3.3 28.7 ⫾ 1.8 17.2 (22/128) 34 (16/47) 25
43 34.5 ⫾ 0.73 11.4 ⫾ 1.0 75 ⫾ 3.1 30.8 ⫾ 2.1 15 (18/120) 30.2 (13/43) 15.4
NS NS NS NS NS NS NS
Conclusions: Implantation and pregnancy rates are slightly higher in the terbutaline group (17.2%, 34%) than in ritodrine group (15%, 30.2%) and control (15.7%, 32%). However, at the present time this differences are not statistically significant.
P-183 Telomerase activity in human blastomeres: a predictor for embryo viability and development? C. Wang, S. Horng, C. Chang, H. Huang, H. Wang, Y. Soong. Chang Gung Memorial Hosp, Kweishan, Taoyuan, Taiwan.
Conclusions: In this program, monochorionic, diamnionic placentation is the most common placentation type observed in monozygotic pregnancies resulting from blastocyst transfers (19/27 patients). According to literature, a double ICM blastocyst should have resulted in a similar monochorionic twin. The occurrance of a dichorionic twin may imply that, in this case, the trophoblast cells were already destined to separate at the time of observation or that the separation could not be observed. Since all of this occurred well before embryo hatching, this case of monozygotic twinning can not be explained by in vitro zona alteration. The occurrance of two ICM’s in the same blastocyst demonstrates at least one likely mechanism by which monozygotic twinning may result when transferring blastocysts. P-182
2 receptor agonists during the peri-implantation period could improve implantation and pregnancy rates. J. G. Franco Jr, R. L. Baruffi, E. Garbellini, V. Felipe, C. Petersen, A. L. Mauri. Ctr for Human Reproduction Sinha´ Junqueira Maternity Fdn, Ribeirao Preto, Brazil. Objective: 2 receptor agonists can decrease uterine activity and increased endometrial blood flow (Akerlund and Andersson, 1976). On the other hand, decreased uterine contractions frequency at the time of embryo transfer is associated with increased implantation and pregnancy rates (Fanchin et al., 1998). Impaired perfusion of endometrial blood flow has an unfavourable reproductive outcome (Yang et al., 1999). The purpose of this study was to compare in a prospective randomized trial the use of two b2 receptor agonists (terbutaline and ritodrine) during the peri-implantation period in terms of the ICSI program outcome. Design: A prospective randomised clinical study. Materials/Methods: A total of 115 patients from ICSI program were random assigned to one of the drugs (2/1) by drawing lots, using a randomization table previously elaborated for the study. The terbutaline group received 10 mg/day for 15 days from the day after oocyte collection up to the pregnancy test (14 days post-embryo transfer). The ritodrine group received 20 mg/day for the same period of time terbutaline group. All women were stimulated for oocyte retrieval with r-FSH (Puregon®) after blockade of as the 2nd phase (starting on the 21st day of the cycle) with nasal nafarelin (Synarel®) at the dose of 400 g a day until the day of administration of human chorionic gonadotropin (hCG). Fixed doses of 150
FERTILITY & STERILITY威
Objective: Telomerase is a ribonucleoprotein that synthesizes telomeric DNA at the ends of eukaryotic chromosomes. It has been hypothesized that telomerase activity is necessary for cellular immortalization and that telomerase activity is present in cells of germline origin and embryos. Consistent with the hypothesis, telomerase activity has been detected in extracts of human ovaries and testes. But it has not been measured in single human blastomere because of difficulty in obtaining sufficient DNA for analysis. Based on the importance of telomerase for meiosis and mitosis, we designed this study for the assessment of telomerase activity in single blastomeres with the attempt to establish a possible selection or scoring system, in order to select a better embryo for uterine transfer, or to be used as a marker for embryonic health. Design: Prospective clinical study with RT-PCR for telomerase activity in individual human blastomeres. Materials/Methods: Oocytes were obtained from patients undergoing an assisted fertilization procedure after ovarian stimulation with GnRH agonist in combination with FSH/ or HMG and hCG. IVF or ICSI were performed as routine. The fertilized oocytes were cultured for three days. Group A consisted of 21 normal cleaving 3PN embryos at 4 to 8 cell stage. Group B consisted of 35 arrested or fragmented embryos at 2 to 4 cell stage, which were considered to be compromised. After removing the zona pellucida. the embryos were placed in calcium- and magnesium-free medium for dissociation of blastomeres. RNA extraction, reverse transcription, and hTCS (human telomerase catalytic subunit) PCR amplication were performed. The amplification resulted in a PCR products size of 145 bp, then were separated by 3% gel electrophoresis. Siha cells and purified water were used as positive and negative control. Results: 95 intact blastomeres were dissociated from 21 embryos in Group A and 110 intact blastomeres from 35 embryos in Group B. Of these, 86 (90.5%) out of 95 blastomeres in Group A and 78 (70.9%) out of 110 blastomeres in Group B demonstrated telomerase activity by the expression of 145 bp product from hTCS mRNA. The difference was significant in statistics (P ⬍ 0.05, Chi-square). Conclusions: The limitations of evaluating embryos based on morphological criteria alone, as are usually done in most infertility centers, are well recognized. Correlations between gross morphology and implantation are weak and inaccurate. Although the results are not satisfactory, many studies have researched more objective criteria for judging embryo viability and implantation potential, in order to select better embryos to transfer, and consequently enhance the implantation and pregnancy rate without increasing the multiple pregnancies. To our knowledge, there is still no report regarding the relationship between telomerase activity and embryo quality. Our study was conducted to assess telomerase activity in individual blas-
S173