Comprehensive chromosome analysis shows a low concordance rate with embryo, blastocoel fluid and spent blastocyst culture medium （第9回 アジア太平洋生殖医学会（ASPIRE））
K. Yoshikai,1 T. Kato2, Y. Matsuda1, M. Kato2, C. Arai1, S. Suzuki1, R. Hana1i, E. Nakano1, Y. Sawada1.3
T. Sawada1, H. Kurahashi2.
1 The Sawada Women’s clinic Fertility Center Nagoya Japan
2 Division of Molecular Genetics, ICMS (Institute for comprehensive Medical Science), Fujita Health University
3 Department of Obstetrics and Gynecology Nagoya City University Hospital
Background and Aims
Using spent blastocyst culture medium or blastocoel fluid (BF) is a noninvasive preimplantation genetic testing compared with Trophectoderm (TE) biopsy.
We performed a chromosome analysis using Next Generation Sequencing (NGS) to compare spent blastocyst culture medium and BF with ICM and TE deriving from the same blastocyst culture.
Material and Method
Vitrified-warmed blastocysts were cultured in 25μl droplets (CSCC with HAS (Irvine Scientific). After 12hours, BF was aspirated from the expanded blastocyst using an ICSI pipette (Oligio). The aspirated liquid was recovered by being discharged directly to 1.5μl PBS placed in the tube. ICM and TE biopsies were performed from re-expanding blastocyst and each sample was placed in the tube containing 1.5μl PBS. The spent culture medium was collected in amounts of 20μl from each sample. All biopsy materials were performed by whole-genome amplification and diagnosed by NGS. Karyotypic evaluation was performed by BlueFuse Multi software.
Result
A total of 45 blastocyst were biopsied that derived from validity data in 31 blastocyst. Amplification rate of BF (63.6%) and spent blastocyst culture medium (46.8%) were brought significantly low compared to ICM (97.8%) and TE (96.9%). Karyotypic concordance rate between BF and ICM was 58.8% and that comparing spent blastocyst culture medium with ICM, its concordance rate was 47.8%. Additionally, BF and spent blastocyst culture medium were many specimens that cannot be determined for karyotype due to high noise.
Conclusion
We can be successful to diagnose by NGS for BF and spent culture medium. A DNA derived from BF and medium are likely to come from apoptotic cells. Therefore, template DNA is fragmented, and its quality is low.
Chromosomal diagnose with BF and spent culture medium has the advantage of low invasiveness, however practical application is difficult because of discordance that it is judged by the fetal karyotype.
T. Sawada1, H. Kurahashi2.
1 The Sawada Women’s clinic Fertility Center Nagoya Japan
2 Division of Molecular Genetics, ICMS (Institute for comprehensive Medical Science), Fujita Health University
3 Department of Obstetrics and Gynecology Nagoya City University Hospital
Background and Aims
Using spent blastocyst culture medium or blastocoel fluid (BF) is a noninvasive preimplantation genetic testing compared with Trophectoderm (TE) biopsy.
We performed a chromosome analysis using Next Generation Sequencing (NGS) to compare spent blastocyst culture medium and BF with ICM and TE deriving from the same blastocyst culture.
Material and Method
Vitrified-warmed blastocysts were cultured in 25μl droplets (CSCC with HAS (Irvine Scientific). After 12hours, BF was aspirated from the expanded blastocyst using an ICSI pipette (Oligio). The aspirated liquid was recovered by being discharged directly to 1.5μl PBS placed in the tube. ICM and TE biopsies were performed from re-expanding blastocyst and each sample was placed in the tube containing 1.5μl PBS. The spent culture medium was collected in amounts of 20μl from each sample. All biopsy materials were performed by whole-genome amplification and diagnosed by NGS. Karyotypic evaluation was performed by BlueFuse Multi software.
Result
A total of 45 blastocyst were biopsied that derived from validity data in 31 blastocyst. Amplification rate of BF (63.6%) and spent blastocyst culture medium (46.8%) were brought significantly low compared to ICM (97.8%) and TE (96.9%). Karyotypic concordance rate between BF and ICM was 58.8% and that comparing spent blastocyst culture medium with ICM, its concordance rate was 47.8%. Additionally, BF and spent blastocyst culture medium were many specimens that cannot be determined for karyotype due to high noise.
Conclusion
We can be successful to diagnose by NGS for BF and spent culture medium. A DNA derived from BF and medium are likely to come from apoptotic cells. Therefore, template DNA is fragmented, and its quality is low.
Chromosomal diagnose with BF and spent culture medium has the advantage of low invasiveness, however practical application is difficult because of discordance that it is judged by the fetal karyotype.
2019.05.08
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