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Is there any difference in vitrification and slow freezing protocol for oocyte cryopreservation?
semen quality; however, it is uncertain if various cancers impact post-thaw sperm survival differently. The objective of this study was to characterize sperm parameters from a thawed semen sample from men with different cancers who cryopreserved prior to onco-therapy. DESIGN: Retrospective review. MATERIALS AND METHODS: We retrospectively evaluated the raw and test thaw semen data from men with newly diagnosed cancer at the University of Washington Male Infertility Laboratory from 1994-2009. A total of 798 semen samples were analyzed from men with the 8 most common cancers of which 477 underwent test thaw. For each raw and test thawed sample sperm concentration and motility were determined and total motile sperm counts (TMC) were calculated. Mean TMCs and changes in TMC for each cancer were compared to normal samples. RESULTS: The most common cancers in this study were: testicular, Hodgkin’s lymphoma, myeloid and lymphoid leukemias, prostate, sarcoma, brain, and lymphocytic cancer NOS. Not surprisingly, healthy patients had the best pre-cryo and post thaw semen quality. Patients with prostate cancer had the best raw and post thaw semen quality (TMC of 155.1 and 53.2, respectively). Lymphoid leukemias demonstrated the worst raw TMC (26.8 M motile/mL), however, myeloid leukemias displayed the worst post thaw TMC (6.9 M motile/mL). All specimens showed severe reductions in TMC after cryopreservation. CONCLUSION: All cryopreserved specimens showed severe decline in the total motile sperm count post thaw. The most severe reduction was seen in myeloid leukemia group, suggesting that these patients should be counseled to provide increased numbers of specimens for fertility preservation.
P-367 Wednesday, October 19, 2011 HIGH IMPLANTATION RATES FOLLOWING SINGLE EMBRYO TRANSFER PROVIDE SIMILAR PREGNANCY RATES TO MULTIPLE EMBRYO TRANSFER USING VITRIFIED DONOR OOCYTES. Z. P. Nagy, C.-C. Chang, D. P. Bernal, A. A. Toledo, D. Mitchell-Leef, D. B. Shapiro. RBA, Reproductive Biology Associates, Sandy Springs, GA. OBJECTIVE: To evaluate the efficiency of single and multiple embryo transfers in a donor-recipient program using vitrified/warmed oocytes. DESIGN: Retrospective study. MATERIALS AND METHODS: A total of 5014 MII oocytes were obtained and vitrified from 225 donors who were screened and tested according to FDA and ASRM guidelines. Eggs were vitrified 3-4 h after retrieval using minimal volume method. ICSI was performed 2-3 h after warming. Embryos were evaluated morphologically prior to transfer, and when high quality embryos were observed the recipient had the possibility of a single embryo transfer. Clinical and laboratory parameters were analyzed using the Oneway ANOVA and the Fisher’s exact test at the level of P<0.05. RESULTS: Donors mean age was 26.1 2.7 years.
No. of recipients Recipient age (mean SD) No. of oocytes warmed (mean SD) No. of oocytes survived (%) No. of oocytes fertilized (%) No. of good quality embryos on Day3 (mean SD) No. of blastocysts (mean SD)* No of embryos for ET (mean SD) No. of embryos re-vitrified (mean SD) No. clinical pregnancies (%) No. of implantations (%)
Single-ET
Multi-ET
P
126 42.2 4.2
351 40.7 4.4
<0.01
711 (5.6 1.8) 2238 (6.4 2.0) <0.01 643 (90.4) 1957 (87.4) 560 (78.8) 1707 (76.3) 380 (3.0 1.6) 1040 (3.0 1.4)
0.04 NS NS
396 (3.5 1.5)
976 (3.6 1.5)
NS
126
725
NA
265 (2.1 1.5) 77 (61.1) 78 (61.9)
356 (1.0 1.3) <0.05 211 (60.1) 317 (43.7)
NS <0.05
There were 105 (49.8%) pregnancies with multiple implantations in the MET group, while there was only one twin pregnancy (due to monozygotic twinning) in the SET group (P<0.001). * Only for Day-5 ET cases (113 for SET and 274 for MET).
FERTILITY & STERILITYÒ
CONCLUSION: The outcome of the current study shows that SET of vitrified/warmed donor oocytes provides both high implantation rates (and virtually no risks for multiples) and high pregnancy rates similar to multiple embryo transfers. Thus, SET should be recommended when high quality embryo(s) are available for qualifying patients to achieve safe pregnancy outcomes.
P-368 Wednesday, October 19, 2011 IS THERE ANY DIFFERENCE IN VITRIFICATION AND SLOW FREEZING PROTOCOL FOR OOCYTE CRYOPRESERVATION? L. Okada, R. Azambuja, V. Lazzari, L. Dorfman, M. Badalotti, A. Petracco. Fertilitat_ Centro de Medicina Reprodutiva, Porto Alegre, RS, Brazil. OBJECTIVE: This study was an observational retrospective cohort, with the objective to compare oocyte cryopreservation efficiency between slow freezing using choline chloride based medium, and vitrification protocol using cryotop. DESIGN: A total of 101 cycles (slow freezing and vitrification techniques) were performed for those patients that decided to thaw their oocytes from 2009 to 2011. MATERIALS AND METHODS: All patients with at least 10 oocytes and not willing to freeze embryos were offered the possibility to cryopreserve oocytes. The oocytes cryopreserved by slow freezing followed Stachecki’s (1998) protocol, while the oocytes vitrified followed Kuwayama’s. (2005) protocol. The endometrium was prepared using Estradiol Benzoate with an initial dose of 2mg/day, starting on the 3rd day of the cycle. The maximum dosage used was 6mg/day when the endometrium reached a thickness of 8mm by ultrasound. Two days before the replacement of the embryos, the oocytes were thawed. After thawing, the oocytes were kept in a incubator during 2 hours before inseminating by ICSI. The survival, fertilization, cleavage, average embryo transfer and pregnancy rates were compared through the chisquare test (P<0.05). RESULTS: The results are as follow: TABLE 1. Outcome results from slow freezing versus vitrification protocol.
Cycles Oocytes Thawed Oocytes Survived* Oocytes Fertilized Embryos Cleaved Embryos Transferred Average Embryo Transfer Implantation Pregnancies Clinical Pregnancies *
Slow freezing (%)
Vitrification (%)
56 523 289 (55.2) 200 (69.2) 185 (92.5) 121 (65.4) 2.2
45 399 255 (63.9) 177 (69.4) 156 (88.1) 100 (64.1) 2.2
15 (12.4) 16 (28.6) 14 (25.0)
16 (16.0) 15 (33.3) 13 (28.9)
P¼0.0085
CONCLUSION: Although the survival rate was statistically higher for the vitrification technique, this did not result in a statistically higher implantation and/or pregnancy rate. Therefore both techniques are reliable to cryopreserve oocytes.
P-369 Wednesday, October 19, 2011 APPLICATION FOR CRYOPRESERVATION OF A SMALL NUMBER OF HUMAN SPERMATOZOA IN A CLOSED SYSTEM BY USING RAPID-IÔ VIA VITRIFICATION. A. Egashira, M. Murakami, K. Tanaka, H. Otsubo, T. Matsuguma, T. Kuramoto. Kuramoto Women’s Clinic, Fukuoka, Japan. OBJECTIVE: In this study, we evaluated the usefulness of the Rapid-iÔ vitrification system for cryopreserving a small number of spermatozoa in a closed system.
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P-367 Wednesday, October 19, 2011 HIGH IMPLANTATION RATES FOLLOWING SINGLE EMBRYO TRANSFER PROVIDE SIMILAR PREGNANCY RATES TO MULTIPLE EMBRYO TRANSFER USING VITRIFIED DONOR OOCYTES. Z. P. Nagy, C.-C. Chang, D. P. Bernal, A. A. Toledo, D. Mitchell-Leef, D. B. Shapiro. RBA, Reproductive Biology Associates, Sandy Springs, GA. OBJECTIVE: To evaluate the efficiency of single and multiple embryo transfers in a donor-recipient program using vitrified/warmed oocytes. DESIGN: Retrospective study. MATERIALS AND METHODS: A total of 5014 MII oocytes were obtained and vitrified from 225 donors who were screened and tested according to FDA and ASRM guidelines. Eggs were vitrified 3-4 h after retrieval using minimal volume method. ICSI was performed 2-3 h after warming. Embryos were evaluated morphologically prior to transfer, and when high quality embryos were observed the recipient had the possibility of a single embryo transfer. Clinical and laboratory parameters were analyzed using the Oneway ANOVA and the Fisher’s exact test at the level of P<0.05. RESULTS: Donors mean age was 26.1 2.7 years.
No. of recipients Recipient age (mean SD) No. of oocytes warmed (mean SD) No. of oocytes survived (%) No. of oocytes fertilized (%) No. of good quality embryos on Day3 (mean SD) No. of blastocysts (mean SD)* No of embryos for ET (mean SD) No. of embryos re-vitrified (mean SD) No. clinical pregnancies (%) No. of implantations (%)
Single-ET
Multi-ET
P
126 42.2 4.2
351 40.7 4.4
<0.01
711 (5.6 1.8) 2238 (6.4 2.0) <0.01 643 (90.4) 1957 (87.4) 560 (78.8) 1707 (76.3) 380 (3.0 1.6) 1040 (3.0 1.4)
0.04 NS NS
396 (3.5 1.5)
976 (3.6 1.5)
NS
126
725
NA
265 (2.1 1.5) 77 (61.1) 78 (61.9)
356 (1.0 1.3) <0.05 211 (60.1) 317 (43.7)
NS <0.05
There were 105 (49.8%) pregnancies with multiple implantations in the MET group, while there was only one twin pregnancy (due to monozygotic twinning) in the SET group (P<0.001). * Only for Day-5 ET cases (113 for SET and 274 for MET).
FERTILITY & STERILITYÒ
CONCLUSION: The outcome of the current study shows that SET of vitrified/warmed donor oocytes provides both high implantation rates (and virtually no risks for multiples) and high pregnancy rates similar to multiple embryo transfers. Thus, SET should be recommended when high quality embryo(s) are available for qualifying patients to achieve safe pregnancy outcomes.
P-368 Wednesday, October 19, 2011 IS THERE ANY DIFFERENCE IN VITRIFICATION AND SLOW FREEZING PROTOCOL FOR OOCYTE CRYOPRESERVATION? L. Okada, R. Azambuja, V. Lazzari, L. Dorfman, M. Badalotti, A. Petracco. Fertilitat_ Centro de Medicina Reprodutiva, Porto Alegre, RS, Brazil. OBJECTIVE: This study was an observational retrospective cohort, with the objective to compare oocyte cryopreservation efficiency between slow freezing using choline chloride based medium, and vitrification protocol using cryotop. DESIGN: A total of 101 cycles (slow freezing and vitrification techniques) were performed for those patients that decided to thaw their oocytes from 2009 to 2011. MATERIALS AND METHODS: All patients with at least 10 oocytes and not willing to freeze embryos were offered the possibility to cryopreserve oocytes. The oocytes cryopreserved by slow freezing followed Stachecki’s (1998) protocol, while the oocytes vitrified followed Kuwayama’s. (2005) protocol. The endometrium was prepared using Estradiol Benzoate with an initial dose of 2mg/day, starting on the 3rd day of the cycle. The maximum dosage used was 6mg/day when the endometrium reached a thickness of 8mm by ultrasound. Two days before the replacement of the embryos, the oocytes were thawed. After thawing, the oocytes were kept in a incubator during 2 hours before inseminating by ICSI. The survival, fertilization, cleavage, average embryo transfer and pregnancy rates were compared through the chisquare test (P<0.05). RESULTS: The results are as follow: TABLE 1. Outcome results from slow freezing versus vitrification protocol.
Cycles Oocytes Thawed Oocytes Survived* Oocytes Fertilized Embryos Cleaved Embryos Transferred Average Embryo Transfer Implantation Pregnancies Clinical Pregnancies *
Slow freezing (%)
Vitrification (%)
56 523 289 (55.2) 200 (69.2) 185 (92.5) 121 (65.4) 2.2
45 399 255 (63.9) 177 (69.4) 156 (88.1) 100 (64.1) 2.2
15 (12.4) 16 (28.6) 14 (25.0)
16 (16.0) 15 (33.3) 13 (28.9)
P¼0.0085
CONCLUSION: Although the survival rate was statistically higher for the vitrification technique, this did not result in a statistically higher implantation and/or pregnancy rate. Therefore both techniques are reliable to cryopreserve oocytes.
P-369 Wednesday, October 19, 2011 APPLICATION FOR CRYOPRESERVATION OF A SMALL NUMBER OF HUMAN SPERMATOZOA IN A CLOSED SYSTEM BY USING RAPID-IÔ VIA VITRIFICATION. A. Egashira, M. Murakami, K. Tanaka, H. Otsubo, T. Matsuguma, T. Kuramoto. Kuramoto Women’s Clinic, Fukuoka, Japan. OBJECTIVE: In this study, we evaluated the usefulness of the Rapid-iÔ vitrification system for cryopreserving a small number of spermatozoa in a closed system.
S215