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Vitrification: what is the outcome using a fda cleared “closed” carrier system?
DESIGN: Retrospective descriptive study of the frequence of donors with cystic fibrosis, fragile X syndrome or abnormal karyotype found out in IVI Seville, between February 2007 and December 2007. MATERIALS AND METHODS: Retrospective descriptive study of the frequence of donors with cystic fibrosis, fragile X syndrome or abnormal karyotype found out in IVI Seville, between February 2007 and December 2007. RESULTS: During this period, 275 volunteered to donate oocytes at our clinic. After we carried out the screening we had to exclude 24 (8,7 %) women for different reasons. Two (9 %)of this 24, were eliminated because of an abnormal karyotype (46XX, 21pþ;46XX, 16phþ) 0,7% of the total, fifteen (62%) were dicarded for presenting cystic fibrosis (twelve in 5T allele and three in A508 allele) 5,4% of the total, and seven (29%) were carriers of fragile X syndrome of permutation in heterozygocity, 2,5% of the total. CONCLUSIONS: Due to the incidence we founds we considered useful to realize these test genetic in all donors before we accept them. This way we decrease the risk of inheritance of a genetic disorder in the children of the oocyte donor program. Supported by: Due to the incidence we founds we considered useful to realize these test genetic in all donors before we accept them. This way we decrease the risk of inheritance of a genetic disorder in the children of the oocyte donor program.
CRYOPRESERVATION P-502 ASSOCIATION OF RE-EXPANSION RATE OF BLASTOCOELE AND PREIMPLANTATION EMBRYO DEVELOPMENT IN RE-FROZEN BLASTOCYSTS. R. Hirata, T. Habara, N. Hayashi, N. Kawakami, N. Yoshioka, Y. Aoi. Okayama Couple’s Clinic, Okayama, Japan. OBJECTIVE: Cryopreservation has become a routine technology in IVF. However, little is known about embryo growth after embryo transfer. The aim of this study was to investigate whether any correlations existed between the re-expansion rate of blastocoele and preimplantation embryo development. DESIGN: Prospective study. MATERIALS AND METHODS: Supernumerary cleavage embryos were cryopreserved with slow freezing. Subsequently, 483 embryos which were requested to be discarded were employed. After thawing, 396 frozen-thawed embryos with less than 50% intact blastomeres were cultured; thereafter, 178 blastocysts were refrozen with vitrification using Cryotop. The recovery rates immediately after re-thawing were classified into 3 groups including less than 80%, 80-100% and more than 100%, while the morphological changes of blastocysts 2 hours and 48 hours after additional incubation (degenerated, recovery of less than 80%, recovery of 80-100%, recovery of more than 100%, hatching and hatched) were compared.Statistical analysis was by Fisher’s exact test or chi-square test with Yates’ correction as appropriate. P values <0.05 were considered significant. The SPSS package was used for the analysis. RESULTS: Of 178 vitrified blastocysts, 175(98.3%) re-expanded. These re-expanded blastocysts were used in the experiment. The percentage of good quality blastocysts was 20%(35/175). Comparison immediately after thawing according to recovery rates revealed no difference in morphological changes in blastocysts 48 hours after thawing. The blastocysts demonstrating higher recovery rates 2 hours after thawing showed a tendency of increase in the number of hatched blastocysts in 48 hours after thawing. The blastocysts which shrank immediately after thawing but had recovered in size 2 hours later showed higher rates of hatched blastocysts. CONCLUSIONS: The results suggested that the re-expansion rate of blastocoele might be a indicator of preimplantation embryo development. Supported by: None.
P-503 VITRIFICATION: WHAT IS THE OUTCOME USING A FDA CLEARED ‘‘CLOSED’’ CARRIER SYSTEM? J. Liebermann, J. M. Matthews, A. J. Erman, S. R. Sanchez, Y. Wagner, E. J. Pelts. Fertility Centers of Illinois, Chicago, IL. OBJECTIVE: The vitrification technique is traditionally based on direct contact between the vitrification solution containing the cryoprotectant
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agents and the liquid nitrogen (LN2), and so the question arises as to whether the LN2 has to be sterilized, as it may be a source of contamination. Most vitrification systems are still using a carrier which require direct contact between the embryos and LN2. Despite the fact that to date no viral, fungal or bacterial contamination event has been described in approximately 1,500 publications related to vitrification since 1985, in August 2007 we switched to an FDA cleared ‘‘closed’’ carrier system and examined these FET’s in terms of their clinical outcome. DESIGN: A retrospective analysis of frozen embryo transfers (FET) using an FDA cleared ‘‘closed’’ carrier system. MATERIALS AND METHODS: A total of 62 FET using the high security vitrification kit (HSV, Cryo Bio System, L’Aigle, France) were included in this study. Both natural and hormone replacement cycles were used to increase the endometrial receptivity, and laboratory protocols were followed, including vitrification and warming of the blastocysts and assisted hatching for all warmed blastocysts except those that were already hatching. The HSV system was used as a carrier, with a mixture of 15% ethylene glycol/DMSO (v/v) þ 0.5M sucrose. RESULTS: In 62 FET, the HSV provided a survival rate in the mid 90% (125/131), and the clinical pregnancy per transfer and implantation rates were 50.0% (31/62) and 33.9% (42/124) respectively.
Patient’s age (y) No. of transfers No. of blastocysts thawed No. of blastocysts survived (%) No. of blastocysts transferred Mean no. of blastocysts transferred No. of implantations (%) No. of positive pregnancy/FET (%) No. of clinical pregnancy/FET (5) Ongoing pregnancies (%)
33.6 3.6 62 131 125 (95.4) 124 2.0 42 (33.9) 37 (59.7) 31 (50.0) 30 (96.8)
CONCLUSIONS: ‘‘Closed’’ vitrification systems are available now to eliminate the potential risk of contaminating human embryos during cryopreservation procedures by direct contact between the sample and LN2. After four years of using an ‘‘open’’ system, our experience with a ‘‘closed’’ system is very positive and promising based on the data outcome. Although high cooling rates are required to achieve a glassy vitrified stage, the lower cooling rates provides by the ‘‘closed’’ system seem suitable to vitrify and warm blastocysts successfully. The HSV kit is now our standard carrier for vitrification of human blastocysts. Supported by: None.
P-504 HUMAN OOCYTE CRYOPRESERVATION: THE OUTCOME OF A NEW OOCYTE FREEZING TECHNIQUE IS COMPARED TO A TRADITIONAL EMBRYO FREEZING PROTOCOL. M. C. Rodriguez-Karl, M. Reynoso, M. Ruiz, J. E. Moody, A.-T. H. La, D. G. Diaz. West Coast Fertility Centers, Fountain Valley, CA. OBJECTIVE: The objective of this study is to compare the outcome of 70 thawed oocytes cycles from a new slow-freeze technique versus thawed embryos using the traditional slow-freeze protocol at a private IVF clinic in a three year interval. DESIGN: This retrospective study examines the thaw survival, pregnancy and an implantation rates between frozen embryo transfer (FET) versus frozen oocytes transfer (FOT), and is being conducted under IRB supervision. MATERIALS AND METHODS: Group A: Patients’ age % 37 with a day 3 serum FSH % 10 underwent IVF and had preservation of surplus embryos (FET). Group B: patients utilized frozen-thawed oocytes (FOT). RESULTS: An analysis was conducted on 214 cycles using frozenthawed human embryos (FET) and on 70 cycles using frozen-thawed human oocytes (FOT). A total of 1096 frozen embryos were thawed and 471 frozen-thawed oocytes. The respective thawed survival rates were 80.5% (882 / 1096) versus 92% (433 / 471), pregnancy rates were 47.2% (101 / 214) versus 54.3% (38 / 70) and implantation was at 16.4% (125 / 763) versus 20.4% (61 / 299).
Vol. 90, Suppl 1, September 2008
CRYOPRESERVATION P-502 ASSOCIATION OF RE-EXPANSION RATE OF BLASTOCOELE AND PREIMPLANTATION EMBRYO DEVELOPMENT IN RE-FROZEN BLASTOCYSTS. R. Hirata, T. Habara, N. Hayashi, N. Kawakami, N. Yoshioka, Y. Aoi. Okayama Couple’s Clinic, Okayama, Japan. OBJECTIVE: Cryopreservation has become a routine technology in IVF. However, little is known about embryo growth after embryo transfer. The aim of this study was to investigate whether any correlations existed between the re-expansion rate of blastocoele and preimplantation embryo development. DESIGN: Prospective study. MATERIALS AND METHODS: Supernumerary cleavage embryos were cryopreserved with slow freezing. Subsequently, 483 embryos which were requested to be discarded were employed. After thawing, 396 frozen-thawed embryos with less than 50% intact blastomeres were cultured; thereafter, 178 blastocysts were refrozen with vitrification using Cryotop. The recovery rates immediately after re-thawing were classified into 3 groups including less than 80%, 80-100% and more than 100%, while the morphological changes of blastocysts 2 hours and 48 hours after additional incubation (degenerated, recovery of less than 80%, recovery of 80-100%, recovery of more than 100%, hatching and hatched) were compared.Statistical analysis was by Fisher’s exact test or chi-square test with Yates’ correction as appropriate. P values <0.05 were considered significant. The SPSS package was used for the analysis. RESULTS: Of 178 vitrified blastocysts, 175(98.3%) re-expanded. These re-expanded blastocysts were used in the experiment. The percentage of good quality blastocysts was 20%(35/175). Comparison immediately after thawing according to recovery rates revealed no difference in morphological changes in blastocysts 48 hours after thawing. The blastocysts demonstrating higher recovery rates 2 hours after thawing showed a tendency of increase in the number of hatched blastocysts in 48 hours after thawing. The blastocysts which shrank immediately after thawing but had recovered in size 2 hours later showed higher rates of hatched blastocysts. CONCLUSIONS: The results suggested that the re-expansion rate of blastocoele might be a indicator of preimplantation embryo development. Supported by: None.
P-503 VITRIFICATION: WHAT IS THE OUTCOME USING A FDA CLEARED ‘‘CLOSED’’ CARRIER SYSTEM? J. Liebermann, J. M. Matthews, A. J. Erman, S. R. Sanchez, Y. Wagner, E. J. Pelts. Fertility Centers of Illinois, Chicago, IL. OBJECTIVE: The vitrification technique is traditionally based on direct contact between the vitrification solution containing the cryoprotectant
S276
Abstracts
agents and the liquid nitrogen (LN2), and so the question arises as to whether the LN2 has to be sterilized, as it may be a source of contamination. Most vitrification systems are still using a carrier which require direct contact between the embryos and LN2. Despite the fact that to date no viral, fungal or bacterial contamination event has been described in approximately 1,500 publications related to vitrification since 1985, in August 2007 we switched to an FDA cleared ‘‘closed’’ carrier system and examined these FET’s in terms of their clinical outcome. DESIGN: A retrospective analysis of frozen embryo transfers (FET) using an FDA cleared ‘‘closed’’ carrier system. MATERIALS AND METHODS: A total of 62 FET using the high security vitrification kit (HSV, Cryo Bio System, L’Aigle, France) were included in this study. Both natural and hormone replacement cycles were used to increase the endometrial receptivity, and laboratory protocols were followed, including vitrification and warming of the blastocysts and assisted hatching for all warmed blastocysts except those that were already hatching. The HSV system was used as a carrier, with a mixture of 15% ethylene glycol/DMSO (v/v) þ 0.5M sucrose. RESULTS: In 62 FET, the HSV provided a survival rate in the mid 90% (125/131), and the clinical pregnancy per transfer and implantation rates were 50.0% (31/62) and 33.9% (42/124) respectively.
Patient’s age (y) No. of transfers No. of blastocysts thawed No. of blastocysts survived (%) No. of blastocysts transferred Mean no. of blastocysts transferred No. of implantations (%) No. of positive pregnancy/FET (%) No. of clinical pregnancy/FET (5) Ongoing pregnancies (%)
33.6 3.6 62 131 125 (95.4) 124 2.0 42 (33.9) 37 (59.7) 31 (50.0) 30 (96.8)
CONCLUSIONS: ‘‘Closed’’ vitrification systems are available now to eliminate the potential risk of contaminating human embryos during cryopreservation procedures by direct contact between the sample and LN2. After four years of using an ‘‘open’’ system, our experience with a ‘‘closed’’ system is very positive and promising based on the data outcome. Although high cooling rates are required to achieve a glassy vitrified stage, the lower cooling rates provides by the ‘‘closed’’ system seem suitable to vitrify and warm blastocysts successfully. The HSV kit is now our standard carrier for vitrification of human blastocysts. Supported by: None.
P-504 HUMAN OOCYTE CRYOPRESERVATION: THE OUTCOME OF A NEW OOCYTE FREEZING TECHNIQUE IS COMPARED TO A TRADITIONAL EMBRYO FREEZING PROTOCOL. M. C. Rodriguez-Karl, M. Reynoso, M. Ruiz, J. E. Moody, A.-T. H. La, D. G. Diaz. West Coast Fertility Centers, Fountain Valley, CA. OBJECTIVE: The objective of this study is to compare the outcome of 70 thawed oocytes cycles from a new slow-freeze technique versus thawed embryos using the traditional slow-freeze protocol at a private IVF clinic in a three year interval. DESIGN: This retrospective study examines the thaw survival, pregnancy and an implantation rates between frozen embryo transfer (FET) versus frozen oocytes transfer (FOT), and is being conducted under IRB supervision. MATERIALS AND METHODS: Group A: Patients’ age % 37 with a day 3 serum FSH % 10 underwent IVF and had preservation of surplus embryos (FET). Group B: patients utilized frozen-thawed oocytes (FOT). RESULTS: An analysis was conducted on 214 cycles using frozenthawed human embryos (FET) and on 70 cycles using frozen-thawed human oocytes (FOT). A total of 1096 frozen embryos were thawed and 471 frozen-thawed oocytes. The respective thawed survival rates were 80.5% (882 / 1096) versus 92% (433 / 471), pregnancy rates were 47.2% (101 / 214) versus 54.3% (38 / 70) and implantation was at 16.4% (125 / 763) versus 20.4% (61 / 299).
Vol. 90, Suppl 1, September 2008