- Email: [email protected]
PGD for cryptic translocation carrier couples
Abstracts – PGDIS: 9th International Symposium on Preimplantation Genetics
PGD for cryptic translocation carrier couples Erçelen N¹, Tutar E¹, Gültomruk M¹, Erkan L¹, Ucar HC¹, Balaban B², Urman B² ¹VKV American Hospital, Genetics and Genomic Sciences Center; ²VKV American Hospital, Women’ s Health Center, Istanbul, Turkey Objective: To present the PGD results of reciprocal and Robertsonian translocations that were analysed in 22 ICSI/ PGD cycles for 18 couples. Materials and methods: Cytogenetic analysis was performed on cultured lymphocytes derived from peripheral bloods of couples. Two telomeric probes with two or one centromeric probes for the chromosomes involved in the translocation were analysed on metaphase spreads of translocation carrier individual. Each blastomere biopsied from day-3 embryos was analysed with telomeric and centromeric probes. They were also analysed for common aneuploidies with PGT probes 13, 18, 21, X, Y (Vysis). Embryos available for transfer were transferred on day 5 of embryo development. Prenatal diagnosis was recommended for pregnant patients.
Results: PGD was performed for 22 translocation carrier cycles. Average maternal age was 32.2 years. Average of 1.6 normal embryos were transferred in 14 cycles. 133 embryos were analysed and only 18% (24/133) were normal or balanced. 16.3% (14/86) of embryos were analysed normal or balanced in reciprocal translocations, while 21.3% (10/47) of embryos were analysed normal or balanced in Robertsonian translocations. Clinical pregnancy per embryo transfer was 20% (2/10) and 50% (2/4) in reciprocal and Robertsonian translocations, respectively. Four clinical pregnancies resulted. Three babies were delivered; one twin pregnancy is still ongoing. Conclusion: Individuals that carry chromosomal translocations seem healthy; however, they can produce chromosomally unbalanced gametes and they are at an increased risk (10– 15%) of a chromosomally abnormal fetus. In conclusion, PGD should be advised for poor-prognosis translocation carrier couples to reduce the number of pregnancy losses and to have successful pregnancy.
Clinical and Social Aspects of PGD Chromosomal abnormalities in preimplantation embryos derived from assisted reproduction techniques Çınar C1, Beyazyurek C1, Onal B1, Ekmekci CG1, Karlikaya G3, Karagozoglu H3, Kavrut M3, Unal S2, Altin G2, Yelke H2, Kilinc C1, Kahraman S3 1Reproductive Genetics Center; 2Embryology Laboratory; 3Assisted Reproduction and In Vitro Fertilization Center, Istanbul Memorial Hospital, Istanbul, Turkey Introduction: PGD has a potential impact on increasing implantation rates and decreasing miscarriages. The purpose of this study is to identify the ratio of chromosomal abnormalities and clinical outcomes in poor-prognosis patients referred for assisted reproduction treatment. Materials and methods: In 2008, 221 PGD cycles were performed mainly for the following indications; advanced maternal age, repeated implantation failures, recurrent pregnancy losses, previous aneuploidic conception and male factor infertility. Nearly half of the cases contained at least one additional PGD indication. Zona drilling was performed by laser and biopsy was done by removal of one blastomere on day 3. Embryo transfers were done mostly on day 4. Blastomeres were fixed using hypotonic/fixative method. FISH for nine chromosomes (13, 15, 16, 17, 18, 21, 22, X, Y) were applied using two consecutive hybridizations, plus rehybridizations using locus-specific and telomeric probes known as no-result rescue when there are doubts in signal evaluations. Multinucleation/quality of nuclei of biopsied blastomere and embryonic developments before and after the biopsy were also assessed. Results: Out of 1738 biopsied embryos, 1538 were analysed by FISH (88.5%). 65.8% of analysed embryos were found to be abnormal. Most of the abnormalities were complex aneuploidies (41.8%) and monosomies (27.4%). Embryo transfer was carried out in 204 cycles, resulting with a clinical pregnancy rate (CPR) of 40.7% and missed abortion rate of 8.4% overall. At the time this report was prepared, 29 healthy babies were born and 42 pregnancies were still continuing. Among 221 cycles, in 94 cycles the age of the woman was
37 years or higher. In this group, the aneuploidy rate (AR) was 74% and CPR was 36.1%. In the young group, AR was 60.9% and CPR was 43.8%. To assess the effect of male factor, young maternal age cases were grouped according to the sperm source; ejaculate versus micro-testicular sperm extraction (TESE). Ejaculate group had 58.8% AR while micro-TESE group had 69.4%. Pregnancy rates were 48.8% and 31.2% for ejaculate and TESE groups, respectively. A tendency to increase in aneuploidy rates and decrease in CPR was observed when additional indications were added. The developmental potentials of normal and abnormal embryos differed significantly. The proportion of slow-growing ones among normal embryos was 17.2%; however, nearly half of the abnormal embryos were developmentally eliminated on day 4 or 5. Nucleus structure/quality was another determining factor for PGD outcomes; multinucleated blastomeres had very low chance of being normal (8.1%) mostly being complex aneuploidic, and most of the embryos containing these types of blastomeres have impaired development (over 60%), indicating unsuitability of multinucleated blastomeres for biopsy and analysis. Conclusion: Although there were limiting factors in PGD cycles such as mosaicism, limited number of chromosomes that were analysed and a pretty high proportion of poorprognosis patients in our series, our results show that acceptable cumulative pregnancies can be achieved even when the combined indications are present. Implications of sperm aneuploidy in PGD Oter M1, Corral S1, Fernández S1, Toro E1, Colomar A1, Casanovas A1, Cañadas C2, Badajoz V2, Verdú V2, LópezTeijón M3, Velilla E1 1Centre for Embryo Medicine; 2GINEFIV; 3Fundación Leonardo Marques, Spain Aim: Couples with sperm aneuploidy are at increased risk of producing aneuploid embryos. Synaptic abnormalities during meiosis can lead to sperm aneuploidy or meiotic arrest. However, depending on the type of meiotic segregation
S-17 Reproductive BioMedicine Online, Vol. 18, Suppl. 3, May 2009
PGD for cryptic translocation carrier couples Erçelen N¹, Tutar E¹, Gültomruk M¹, Erkan L¹, Ucar HC¹, Balaban B², Urman B² ¹VKV American Hospital, Genetics and Genomic Sciences Center; ²VKV American Hospital, Women’ s Health Center, Istanbul, Turkey Objective: To present the PGD results of reciprocal and Robertsonian translocations that were analysed in 22 ICSI/ PGD cycles for 18 couples. Materials and methods: Cytogenetic analysis was performed on cultured lymphocytes derived from peripheral bloods of couples. Two telomeric probes with two or one centromeric probes for the chromosomes involved in the translocation were analysed on metaphase spreads of translocation carrier individual. Each blastomere biopsied from day-3 embryos was analysed with telomeric and centromeric probes. They were also analysed for common aneuploidies with PGT probes 13, 18, 21, X, Y (Vysis). Embryos available for transfer were transferred on day 5 of embryo development. Prenatal diagnosis was recommended for pregnant patients.
Results: PGD was performed for 22 translocation carrier cycles. Average maternal age was 32.2 years. Average of 1.6 normal embryos were transferred in 14 cycles. 133 embryos were analysed and only 18% (24/133) were normal or balanced. 16.3% (14/86) of embryos were analysed normal or balanced in reciprocal translocations, while 21.3% (10/47) of embryos were analysed normal or balanced in Robertsonian translocations. Clinical pregnancy per embryo transfer was 20% (2/10) and 50% (2/4) in reciprocal and Robertsonian translocations, respectively. Four clinical pregnancies resulted. Three babies were delivered; one twin pregnancy is still ongoing. Conclusion: Individuals that carry chromosomal translocations seem healthy; however, they can produce chromosomally unbalanced gametes and they are at an increased risk (10– 15%) of a chromosomally abnormal fetus. In conclusion, PGD should be advised for poor-prognosis translocation carrier couples to reduce the number of pregnancy losses and to have successful pregnancy.
Clinical and Social Aspects of PGD Chromosomal abnormalities in preimplantation embryos derived from assisted reproduction techniques Çınar C1, Beyazyurek C1, Onal B1, Ekmekci CG1, Karlikaya G3, Karagozoglu H3, Kavrut M3, Unal S2, Altin G2, Yelke H2, Kilinc C1, Kahraman S3 1Reproductive Genetics Center; 2Embryology Laboratory; 3Assisted Reproduction and In Vitro Fertilization Center, Istanbul Memorial Hospital, Istanbul, Turkey Introduction: PGD has a potential impact on increasing implantation rates and decreasing miscarriages. The purpose of this study is to identify the ratio of chromosomal abnormalities and clinical outcomes in poor-prognosis patients referred for assisted reproduction treatment. Materials and methods: In 2008, 221 PGD cycles were performed mainly for the following indications; advanced maternal age, repeated implantation failures, recurrent pregnancy losses, previous aneuploidic conception and male factor infertility. Nearly half of the cases contained at least one additional PGD indication. Zona drilling was performed by laser and biopsy was done by removal of one blastomere on day 3. Embryo transfers were done mostly on day 4. Blastomeres were fixed using hypotonic/fixative method. FISH for nine chromosomes (13, 15, 16, 17, 18, 21, 22, X, Y) were applied using two consecutive hybridizations, plus rehybridizations using locus-specific and telomeric probes known as no-result rescue when there are doubts in signal evaluations. Multinucleation/quality of nuclei of biopsied blastomere and embryonic developments before and after the biopsy were also assessed. Results: Out of 1738 biopsied embryos, 1538 were analysed by FISH (88.5%). 65.8% of analysed embryos were found to be abnormal. Most of the abnormalities were complex aneuploidies (41.8%) and monosomies (27.4%). Embryo transfer was carried out in 204 cycles, resulting with a clinical pregnancy rate (CPR) of 40.7% and missed abortion rate of 8.4% overall. At the time this report was prepared, 29 healthy babies were born and 42 pregnancies were still continuing. Among 221 cycles, in 94 cycles the age of the woman was
37 years or higher. In this group, the aneuploidy rate (AR) was 74% and CPR was 36.1%. In the young group, AR was 60.9% and CPR was 43.8%. To assess the effect of male factor, young maternal age cases were grouped according to the sperm source; ejaculate versus micro-testicular sperm extraction (TESE). Ejaculate group had 58.8% AR while micro-TESE group had 69.4%. Pregnancy rates were 48.8% and 31.2% for ejaculate and TESE groups, respectively. A tendency to increase in aneuploidy rates and decrease in CPR was observed when additional indications were added. The developmental potentials of normal and abnormal embryos differed significantly. The proportion of slow-growing ones among normal embryos was 17.2%; however, nearly half of the abnormal embryos were developmentally eliminated on day 4 or 5. Nucleus structure/quality was another determining factor for PGD outcomes; multinucleated blastomeres had very low chance of being normal (8.1%) mostly being complex aneuploidic, and most of the embryos containing these types of blastomeres have impaired development (over 60%), indicating unsuitability of multinucleated blastomeres for biopsy and analysis. Conclusion: Although there were limiting factors in PGD cycles such as mosaicism, limited number of chromosomes that were analysed and a pretty high proportion of poorprognosis patients in our series, our results show that acceptable cumulative pregnancies can be achieved even when the combined indications are present. Implications of sperm aneuploidy in PGD Oter M1, Corral S1, Fernández S1, Toro E1, Colomar A1, Casanovas A1, Cañadas C2, Badajoz V2, Verdú V2, LópezTeijón M3, Velilla E1 1Centre for Embryo Medicine; 2GINEFIV; 3Fundación Leonardo Marques, Spain Aim: Couples with sperm aneuploidy are at increased risk of producing aneuploid embryos. Synaptic abnormalities during meiosis can lead to sperm aneuploidy or meiotic arrest. However, depending on the type of meiotic segregation
S-17 Reproductive BioMedicine Online, Vol. 18, Suppl. 3, May 2009