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Correlation between prognostic chromosomal segregation and detected rearrangements in preimplantation embryos in a balanced translocation carrier
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RBMO VOLUME 38 ISSUE S1 2019
immediately reverse transcribed to cDNA. In the second subgroup, PCR was performed directly on the culture medium. PCR was performed for SRY and GAPDH genes. The SRY positive embryos were considered as male embryos and those were GAPDH positive and SRY negative considered as females. The results of sexing based on spent culture media was compared with the results of sexing based on FISH. Results: For the first group, 12 samples were evaluated, after comparing the results of PCR and FISH, the results for all the samples were concordant.In the second group, we were able to correctly diagnose 12 of the 14 samples. Two embryos with false diagnosis were females that diagnosed as males by direct PCR. Conclusion: Preimplantation sexing without embryo biopsy by RTPCR on the spent embryo culture media seems to be a reliable tool for non-invasive preimplantation sexing. However, direct PCR-based diagnosis might be lead to false results; this is probably due to DNA contamination. Keywords: Preimplantation, Sexing, Culture medium
doi: 10.1016/j.rbmo.2019.03.059
13. CORRELATION BETWEEN PROGNOSTIC CHROMOSOMAL SEGREGATION AND DETECTED REARRANGEMENTS IN PREIMPLANTATION EMBRYOS IN A BALANCED TRANSLOCATION CARRIER
E.V. Marinova1, M.N. Popova2, D.A. Tzoneva2, I.A. Antonova2, D.M. Gulenova2, G.I. Nikolov2, V.M. Penchev3 1
Office for Technology Transfer in the area of Molecular Genetics and Diagnostics(OTTMGD), Medical Centre “ReproBioMed“ Ltd., Sofia, Bulgaria 2 Department of Assisted Reproduction, Medical Centre “ReproBioMed“ Ltd., Sofia, Bulgaria 3 Department of Assisted Reproduction, Medical Centre “Adella Fertility“ JSC., Sofia, Bulgaria
Introduction: Individuals with balanced translocations are estimated to be around 1 % of the patients with infertility. A reciprocal translocation usually involves breakage of two non-homologous chromosomes with exchange of fragment with different size. The medical history of affected couples includes abortions, stillbirth, congenital anomalies, possible mental retardation and unsuccessful IVF attempts. It is hard to define the exact risk of unbalanced karyotypes in gametes, but it is accepted that it could be more than 80%. Preimplantation genetic screening (PGS) is strongly recommended in cases with chromosome rearrangements allowing the study of female gamete segregation. Material and Methods: In the present case we performed two IVF-PGS cycles in a couple with a 34 years old woman with karyotype 46 XX, t(1;16)(q12;q13). Altogether, 14 embryos on day 3 where biopsied and tested using the 24sure (Illumina Inc., USA) protocol for BAC arrays. Results: Out of the 14 tested embryos only two were euploid, five embryos had unbalanced genome due to the involvement of the chromosomes with the parental translocation. The other embryos had ‘sporadic'aneuploidy. After the PGS an embryotransfer of two euploid embryos was performed with one healthy baby born. Discussion: Commonly more than 60 % of embryos obtained from patients with reciprocal translocations carry abnormal chromosomal constitution and about 1:28/30 are normal. In this case the percentage of unbalanced embryos - 5/14 (∼ 36 %) was significantly lower than the expected according to the published data. However, other independent aneuploidies also co-exist, most of which are age dependent. Conclusions: The theoretical percentage of embryos with
unbalanced karyotypes and the data from the clinical practice after performing PGS in IVF patients with balanced translocations does not match in every case. The phenomenon reveals the need of additional studies about the molecular mechanisms of chromosomal segregation in human gametes and preimplantation embryos delivered from balanced translocation carriers. Keywords: PGD, Balanced translocation doi: 10.1016/j.rbmo.2019.03.060
14. METFORMIN DECREASES THE ANEUPLOIDY RATE IN THE HUMAN PREIMPLANTATION EMBRYOS OF THE PATIENTS WITH POLYCYSTIC OVARIAN SYNDROME
Niloufar Jafarian-Motlagh1,2, Masood Bazrgar2, Hamid Gourabi2, Bahar Movaghar3, Mostafa Fakhri2 1
Department of Genetics, University of Science and Culture, ACECR, Tehran, Iran 2 Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran 3 Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Background: Diploid-aneuploid mosaicism is prevalent in preimplantation stage while transfer of such embryos in case of having enough normal cells could lead to live birth. The results of Chromosomal analysis of blastocyst cultured in diabetic conditions represents that these conditions could lead to increase the number of aneuploid cells. Metformin is a drug that widely prescribed for type 2 diabetes (T2D) and polycystic ovarian syndrome (PCOS). Purpose of this study was evaluation the effect of metformin on aneuploidy rate of preimplantation human embryos. Methods: We randomly included spare embryos of infertile couples with histories of PCOS and male infertility in two groups: treatment with metformin (exposed) and
RBMO VOLUME 38 ISSUE S1 2019
immediately reverse transcribed to cDNA. In the second subgroup, PCR was performed directly on the culture medium. PCR was performed for SRY and GAPDH genes. The SRY positive embryos were considered as male embryos and those were GAPDH positive and SRY negative considered as females. The results of sexing based on spent culture media was compared with the results of sexing based on FISH. Results: For the first group, 12 samples were evaluated, after comparing the results of PCR and FISH, the results for all the samples were concordant.In the second group, we were able to correctly diagnose 12 of the 14 samples. Two embryos with false diagnosis were females that diagnosed as males by direct PCR. Conclusion: Preimplantation sexing without embryo biopsy by RTPCR on the spent embryo culture media seems to be a reliable tool for non-invasive preimplantation sexing. However, direct PCR-based diagnosis might be lead to false results; this is probably due to DNA contamination. Keywords: Preimplantation, Sexing, Culture medium
doi: 10.1016/j.rbmo.2019.03.059
13. CORRELATION BETWEEN PROGNOSTIC CHROMOSOMAL SEGREGATION AND DETECTED REARRANGEMENTS IN PREIMPLANTATION EMBRYOS IN A BALANCED TRANSLOCATION CARRIER
E.V. Marinova1, M.N. Popova2, D.A. Tzoneva2, I.A. Antonova2, D.M. Gulenova2, G.I. Nikolov2, V.M. Penchev3 1
Office for Technology Transfer in the area of Molecular Genetics and Diagnostics(OTTMGD), Medical Centre “ReproBioMed“ Ltd., Sofia, Bulgaria 2 Department of Assisted Reproduction, Medical Centre “ReproBioMed“ Ltd., Sofia, Bulgaria 3 Department of Assisted Reproduction, Medical Centre “Adella Fertility“ JSC., Sofia, Bulgaria
Introduction: Individuals with balanced translocations are estimated to be around 1 % of the patients with infertility. A reciprocal translocation usually involves breakage of two non-homologous chromosomes with exchange of fragment with different size. The medical history of affected couples includes abortions, stillbirth, congenital anomalies, possible mental retardation and unsuccessful IVF attempts. It is hard to define the exact risk of unbalanced karyotypes in gametes, but it is accepted that it could be more than 80%. Preimplantation genetic screening (PGS) is strongly recommended in cases with chromosome rearrangements allowing the study of female gamete segregation. Material and Methods: In the present case we performed two IVF-PGS cycles in a couple with a 34 years old woman with karyotype 46 XX, t(1;16)(q12;q13). Altogether, 14 embryos on day 3 where biopsied and tested using the 24sure (Illumina Inc., USA) protocol for BAC arrays. Results: Out of the 14 tested embryos only two were euploid, five embryos had unbalanced genome due to the involvement of the chromosomes with the parental translocation. The other embryos had ‘sporadic'aneuploidy. After the PGS an embryotransfer of two euploid embryos was performed with one healthy baby born. Discussion: Commonly more than 60 % of embryos obtained from patients with reciprocal translocations carry abnormal chromosomal constitution and about 1:28/30 are normal. In this case the percentage of unbalanced embryos - 5/14 (∼ 36 %) was significantly lower than the expected according to the published data. However, other independent aneuploidies also co-exist, most of which are age dependent. Conclusions: The theoretical percentage of embryos with
unbalanced karyotypes and the data from the clinical practice after performing PGS in IVF patients with balanced translocations does not match in every case. The phenomenon reveals the need of additional studies about the molecular mechanisms of chromosomal segregation in human gametes and preimplantation embryos delivered from balanced translocation carriers. Keywords: PGD, Balanced translocation doi: 10.1016/j.rbmo.2019.03.060
14. METFORMIN DECREASES THE ANEUPLOIDY RATE IN THE HUMAN PREIMPLANTATION EMBRYOS OF THE PATIENTS WITH POLYCYSTIC OVARIAN SYNDROME
Niloufar Jafarian-Motlagh1,2, Masood Bazrgar2, Hamid Gourabi2, Bahar Movaghar3, Mostafa Fakhri2 1
Department of Genetics, University of Science and Culture, ACECR, Tehran, Iran 2 Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran 3 Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Background: Diploid-aneuploid mosaicism is prevalent in preimplantation stage while transfer of such embryos in case of having enough normal cells could lead to live birth. The results of Chromosomal analysis of blastocyst cultured in diabetic conditions represents that these conditions could lead to increase the number of aneuploid cells. Metformin is a drug that widely prescribed for type 2 diabetes (T2D) and polycystic ovarian syndrome (PCOS). Purpose of this study was evaluation the effect of metformin on aneuploidy rate of preimplantation human embryos. Methods: We randomly included spare embryos of infertile couples with histories of PCOS and male infertility in two groups: treatment with metformin (exposed) and