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Transfer and uterine factors are the major recipient-related predictors of success with donor eggs
mental status. Moreover, there is a suggestion that MPA may enhance energy level in comparison to micronized progesterone. Of equal importance is our finding that neither regimen was associated with notable untoward effects on subjective physical findings and on two important dimensions, mental sharpness and sound sleep, either regimen was associated with an improvement in self-reported functioning. This suggests there is an ongoing need for further studies of the psychosocial impact of hormone therapy.
O-9 Transfer and uterine factors are the major recipient-related predictors of success with donor eggs. U. Zenke, R.J. Chetkowski, A. Bates. In Vitro Fertilization Program, Berkeley, CA. Background: Ovum donation has the highest success rate of all assisted reproductive technologies, yet only 41.6% of the 5,844 fresh donor egg transfers reported to the CDC in 1999 resulted in a live birth. The causes of failure have proven difficult to study because of the heterogeneity of donors, recipients, and programs. Objective: To define recipient-related variables affecting the outcome of donor egg cycles by keeping the oocyte and laboratory components constant. Materials and Methods: The subjects consisted of 116 infertile women who underwent 134 donor egg cycles in which two recipients shared eggs from a single, young, fertile, compensated, anonymous donor. The variables examined included: age, body mass index, gravidity, preexisting medical conditions, endometriosis, uterine pathology, endometrial thickness, male factor, the use of ICSI, number of eggs, fertilization rate, number of embryos, embryo quality, number of fresh embryos per transfer, number of embryos frozen, difficulty of transfer, and the ongoing pregnancy rate. Results: Out of the 67 recipient pairs, the subgroup of 41 pairs with discordant outcomes was the primary focus of analysis. Within this subgroup, non-pregnant subjects had higher frequency (31.7% v 9.7%, P ⫽ 0.04) and greater severity of uterine pathology than the pregnant controls. Non-pregnant women were more likely to have either moderate or difficult embryo transfers (31.7% v 9.7%, P ⫽ 0.04). Whereas the mean endometrial thickness did not differ, 6 of 41 non-pregnant patients (14.5%), but none of the pregnant subjects, had endometrium ⬍ 8 mm (P ⫽ 0.01). In the non-pregnant group, at least one of the above three factors was present in 56.1% of cycles compared to only 19.5% of transfers in the pregnant group (P ⫽ 0.001). None of the remaining variables differed between the groups. Conclusion: Analysis of discordant recipient pairs permits the definition of recipient-related determinants of success by assuring that quality of the eggs and of the laboratory are both constant and adequate for conception. Uterine pathology, embryo transfer, and thin endometrium appear to be the main recipient-related variables resulting in failure to establish pregnancy with donor eggs.
O-10 In vitro maturation of fresh and frozen-thawed testicular sperm to optimized ICSI in azoospermic patients. B. Wu, D. Wong, S. Dickstein, T.J. Gelety. Arizona Center for Reproductive Endocrinology and Infertility, Tucson, AZ. Background: Surgical retrieval of testicular sperm for intracytoplasmic sperm injection (ICSI) is now a widely practiced technique in the treatment of azoospermic males. However, testicular biopsy is an invasive procedure. Optimal management of testicular biopsies would therefore reduce surgical extraction to the retrieval of a single sample. Objective: The objective of this study is to evaluate the optimal use of fresh and frozen-thawed testicular biopsy specimens from patients with obstructive and non-obstructive azoospermia. Materials and Methods: Twenty patients suffering from obstructive or non-obstructive azoospermia underwent testicular sperm extraction (TESE) for ICSI cycles between January 2001 and August 2002. The biopsies were obtained during open surgery under local anesthetic. After obtaining testicular tissue, examination for motile sperm was immediately performed. The TESE samples were washed with modified HTF medium and divided into two parts. The first part consisting of a fresh sample was cultured for 1–5 days and motile sperm was utilized for ICSI. The second part was frozen
FERTILITY & STERILITY威
and kept in liquid nitrogen for future use. A total of 10 fresh TESE cycles and 17 frozen TESE cycles were used for ICSI. After ICSI, the fertilization rate (2PN) and embryo quality were recorded. Before ICSI, fresh or frozen TESE specimens were cultured in 37° C for a period of 1 to 5 days and observed for motile sperm including tail twitching. Results: A total of 10 fresh and 17 frozen ICSI cycles were evaluated utilizing TESE sperm. In the 10 fresh TESE ICSI cycles, a total of 149 oocytes were injected using motile sperm (swimming or twitching) and 113 oocytes exemplified normal 2PN fertilized embryos (fertilization rate, 75.8%). In 17 frozen ICSI cycles, a total of 186 oocytes were injected using motile sperm and 139 oocytes showed a 2PN fertilization rate of 74.7%. Comparing Day 3 embryo quality, the best embryos with no fragmentation were seen in 73% of fresh TESE cycles and 65% of frozen TESE cycles (p ⬎ 0.05). To optimize motile TESE sperm for the use in ICSI, the fresh and frozen TESE specimens were cultured in vitro for up to 5 days. The sperm survival rate was summarized in the following table. Cultured Days Fresh TESE Frozen TESE
0
1
3.10% 8.50% 2.50% 10.20%
2 20.50% 20.80%
3
4
18.50% 9.20% 21.50% 10.50%
5 5.00% 4.30%
These results indicate no significant difference in pre- or post-frozen TESE sperm survival. On the first day after testicular sperm extraction, very few motile sperm were observed. After a 24-hour culture, a marked increase in motile sperm is seen with a maximum motility rate between 48 and 72 hours of culture. Conclusions: The data suggest that the freezing and culturing of testicular biopsy specimens is a very reliable approach for assisted reproductive technologies. It offers the possibility of several attempts at IVF/ICSI from a single sample. Two to three days of culture before an oocyte retrieval may increase the availability of motile spermatozoa for intracytoplasmic injection.
O-11 Effect of endogenous and exogenous estrogens on serum levels of vascular cell adhesion molecule-1 (sVCAM-1). I. Souter,1 C. Janzen,1 O. Martinez-Maza,1 F. Stanczyk,2 G. Chaudhuri,1 L. Nathan1. 1Departments of Ob/Gyn, David Geffen School of Medicine at UCLA and 2University of Southern California Keck School of Medicine, Los Angeles, CA. Background: Estradiol may down regulate the expression of VCAM-1, an adhesion molecule implicated in the initiation of atherogenesis. Objective: To assess whether: a) short term changes in estradiol during the different phases of the menstrual cycle alter serum levels of sVCAM-1, b) VCAM-1 expression is suppressed in long term users of exogenous estrogens and c) natural and surgical menopause have an effect on circulating levels of VCAM-1. Materials and Methods: Serum was obtained from the following groups of patients: Group A: 31 normally menstruating women. Serum was obtained on days 1, 12, and 21 of the menstrual cycle. Group B: long term users of exogenous estrogens-37 women receiving oral contraceptive pills (OCP’s) and 42 postmenopausal women on hormone replacement therapy (HRT). Serum was obtained once. Group C: 31 postmenopausal women not receiving HRT. Serum was obtained once. Group D: 7 women undergoing bilateral salpingoophorectomy for benign disease. Serum was obtained twice: prior to and 4 weeks after the BSO. All samples were assayed for 17-estradiol and sVCAM-1 (RIA and ELISA respectively). Results: No significant change was noted in the mean values of sVCAM-1 throughout the various phases of the menstrual cycle, despite a significant change in 17-estradiol (day-1: 566 ⫾ 133 ng/mL, day-12: 562 ⫾ 123 ng/mL, day-21: 554 ⫾ 128 ng/mL). Postmenopausal women receiving HRT had significantly lower levels of sVCAM-1 compared to the ones not receiving HRT (448.8 ⫾ 95 vs 509.5 ⫾ 136 ng/mL, p:0.037). Long term users of OCP’s had significantly lower levels of sVCAM-1 when compared to normally menstruating women throughout the various phases of the cycle (453 ⫾ 93 ng/mL, p ⬍ 0.0003). Four weeks post BSO, patients had increased levels of sVCAM-1 compared to menstruating women and HRT users (771 ⫾ 215 ng/mL, p:0.05 and p:0.005 respectively).
S9
O-9 Transfer and uterine factors are the major recipient-related predictors of success with donor eggs. U. Zenke, R.J. Chetkowski, A. Bates. In Vitro Fertilization Program, Berkeley, CA. Background: Ovum donation has the highest success rate of all assisted reproductive technologies, yet only 41.6% of the 5,844 fresh donor egg transfers reported to the CDC in 1999 resulted in a live birth. The causes of failure have proven difficult to study because of the heterogeneity of donors, recipients, and programs. Objective: To define recipient-related variables affecting the outcome of donor egg cycles by keeping the oocyte and laboratory components constant. Materials and Methods: The subjects consisted of 116 infertile women who underwent 134 donor egg cycles in which two recipients shared eggs from a single, young, fertile, compensated, anonymous donor. The variables examined included: age, body mass index, gravidity, preexisting medical conditions, endometriosis, uterine pathology, endometrial thickness, male factor, the use of ICSI, number of eggs, fertilization rate, number of embryos, embryo quality, number of fresh embryos per transfer, number of embryos frozen, difficulty of transfer, and the ongoing pregnancy rate. Results: Out of the 67 recipient pairs, the subgroup of 41 pairs with discordant outcomes was the primary focus of analysis. Within this subgroup, non-pregnant subjects had higher frequency (31.7% v 9.7%, P ⫽ 0.04) and greater severity of uterine pathology than the pregnant controls. Non-pregnant women were more likely to have either moderate or difficult embryo transfers (31.7% v 9.7%, P ⫽ 0.04). Whereas the mean endometrial thickness did not differ, 6 of 41 non-pregnant patients (14.5%), but none of the pregnant subjects, had endometrium ⬍ 8 mm (P ⫽ 0.01). In the non-pregnant group, at least one of the above three factors was present in 56.1% of cycles compared to only 19.5% of transfers in the pregnant group (P ⫽ 0.001). None of the remaining variables differed between the groups. Conclusion: Analysis of discordant recipient pairs permits the definition of recipient-related determinants of success by assuring that quality of the eggs and of the laboratory are both constant and adequate for conception. Uterine pathology, embryo transfer, and thin endometrium appear to be the main recipient-related variables resulting in failure to establish pregnancy with donor eggs.
O-10 In vitro maturation of fresh and frozen-thawed testicular sperm to optimized ICSI in azoospermic patients. B. Wu, D. Wong, S. Dickstein, T.J. Gelety. Arizona Center for Reproductive Endocrinology and Infertility, Tucson, AZ. Background: Surgical retrieval of testicular sperm for intracytoplasmic sperm injection (ICSI) is now a widely practiced technique in the treatment of azoospermic males. However, testicular biopsy is an invasive procedure. Optimal management of testicular biopsies would therefore reduce surgical extraction to the retrieval of a single sample. Objective: The objective of this study is to evaluate the optimal use of fresh and frozen-thawed testicular biopsy specimens from patients with obstructive and non-obstructive azoospermia. Materials and Methods: Twenty patients suffering from obstructive or non-obstructive azoospermia underwent testicular sperm extraction (TESE) for ICSI cycles between January 2001 and August 2002. The biopsies were obtained during open surgery under local anesthetic. After obtaining testicular tissue, examination for motile sperm was immediately performed. The TESE samples were washed with modified HTF medium and divided into two parts. The first part consisting of a fresh sample was cultured for 1–5 days and motile sperm was utilized for ICSI. The second part was frozen
FERTILITY & STERILITY威
and kept in liquid nitrogen for future use. A total of 10 fresh TESE cycles and 17 frozen TESE cycles were used for ICSI. After ICSI, the fertilization rate (2PN) and embryo quality were recorded. Before ICSI, fresh or frozen TESE specimens were cultured in 37° C for a period of 1 to 5 days and observed for motile sperm including tail twitching. Results: A total of 10 fresh and 17 frozen ICSI cycles were evaluated utilizing TESE sperm. In the 10 fresh TESE ICSI cycles, a total of 149 oocytes were injected using motile sperm (swimming or twitching) and 113 oocytes exemplified normal 2PN fertilized embryos (fertilization rate, 75.8%). In 17 frozen ICSI cycles, a total of 186 oocytes were injected using motile sperm and 139 oocytes showed a 2PN fertilization rate of 74.7%. Comparing Day 3 embryo quality, the best embryos with no fragmentation were seen in 73% of fresh TESE cycles and 65% of frozen TESE cycles (p ⬎ 0.05). To optimize motile TESE sperm for the use in ICSI, the fresh and frozen TESE specimens were cultured in vitro for up to 5 days. The sperm survival rate was summarized in the following table. Cultured Days Fresh TESE Frozen TESE
0
1
3.10% 8.50% 2.50% 10.20%
2 20.50% 20.80%
3
4
18.50% 9.20% 21.50% 10.50%
5 5.00% 4.30%
These results indicate no significant difference in pre- or post-frozen TESE sperm survival. On the first day after testicular sperm extraction, very few motile sperm were observed. After a 24-hour culture, a marked increase in motile sperm is seen with a maximum motility rate between 48 and 72 hours of culture. Conclusions: The data suggest that the freezing and culturing of testicular biopsy specimens is a very reliable approach for assisted reproductive technologies. It offers the possibility of several attempts at IVF/ICSI from a single sample. Two to three days of culture before an oocyte retrieval may increase the availability of motile spermatozoa for intracytoplasmic injection.
O-11 Effect of endogenous and exogenous estrogens on serum levels of vascular cell adhesion molecule-1 (sVCAM-1). I. Souter,1 C. Janzen,1 O. Martinez-Maza,1 F. Stanczyk,2 G. Chaudhuri,1 L. Nathan1. 1Departments of Ob/Gyn, David Geffen School of Medicine at UCLA and 2University of Southern California Keck School of Medicine, Los Angeles, CA. Background: Estradiol may down regulate the expression of VCAM-1, an adhesion molecule implicated in the initiation of atherogenesis. Objective: To assess whether: a) short term changes in estradiol during the different phases of the menstrual cycle alter serum levels of sVCAM-1, b) VCAM-1 expression is suppressed in long term users of exogenous estrogens and c) natural and surgical menopause have an effect on circulating levels of VCAM-1. Materials and Methods: Serum was obtained from the following groups of patients: Group A: 31 normally menstruating women. Serum was obtained on days 1, 12, and 21 of the menstrual cycle. Group B: long term users of exogenous estrogens-37 women receiving oral contraceptive pills (OCP’s) and 42 postmenopausal women on hormone replacement therapy (HRT). Serum was obtained once. Group C: 31 postmenopausal women not receiving HRT. Serum was obtained once. Group D: 7 women undergoing bilateral salpingoophorectomy for benign disease. Serum was obtained twice: prior to and 4 weeks after the BSO. All samples were assayed for 17-estradiol and sVCAM-1 (RIA and ELISA respectively). Results: No significant change was noted in the mean values of sVCAM-1 throughout the various phases of the menstrual cycle, despite a significant change in 17-estradiol (day-1: 566 ⫾ 133 ng/mL, day-12: 562 ⫾ 123 ng/mL, day-21: 554 ⫾ 128 ng/mL). Postmenopausal women receiving HRT had significantly lower levels of sVCAM-1 compared to the ones not receiving HRT (448.8 ⫾ 95 vs 509.5 ⫾ 136 ng/mL, p:0.037). Long term users of OCP’s had significantly lower levels of sVCAM-1 when compared to normally menstruating women throughout the various phases of the cycle (453 ⫾ 93 ng/mL, p ⬍ 0.0003). Four weeks post BSO, patients had increased levels of sVCAM-1 compared to menstruating women and HRT users (771 ⫾ 215 ng/mL, p:0.05 and p:0.005 respectively).
S9