Skip to main content
Top
Published in: Journal of Assisted Reproduction and Genetics 4/2018

01-04-2018 | Embryo Biology

Blastulation timing is associated with differential mitochondrial content in euploid embryos

Authors: Jacqueline R. Ho, Nabil Arrach, Katherine Rhodes-Long, Wael Salem, Lynda K. McGinnis, Karine Chung, Kristin A. Bendikson, Richard J. Paulson, Ali Ahmady

Published in: Journal of Assisted Reproduction and Genetics | Issue 4/2018

Login to get access

Abstract

Purpose

Preimplantation genetic screening (PGS) and assessment of mitochondrial content (MC) are current methods for selection of the best embryos for transfer. Studies suggest that time-lapse morphokinetics (TLM) may also be helpful for selecting embryos more likely to implant. In our study, we sought to examine the relationship between TLM parameters and MC to determine if they could be used adjunctively in embryo selection. We also examined the relationship between MC with ploidy and blastulation.

Methods

Cryopreserved human embryos at the zygote stage were thawed and cultured in a time-lapse system. Blastomere and trophectoderm biopsies were performed on days 3 and 6. Biopsied cells and all whole embryos from day 6 were analyzed for MC (ratio of mitochondrial to nuclear DNA) and ploidy using next-generation sequencing.

Results

In embryos, MC per cell declined between day 3 and day 6. While early cleavage parameters did not predict MC, embryos with longer blastulation timing had higher MC on day 6. Day 6 MC was lower in euploid vs. aneuploid embryos and lower in blastocysts vs. arrested embryos.

Conclusions

A lower MC at the blastocyst stage was associated with euploid status and blastocyst formation, indicating better embryo quality compared to those with a higher MC. Higher MC in aneuploid and arrested embryos may be explained by slower cell division or degradation of genomic DNA over time. Blastulation timing may be helpful for selection of higher quality embryos. Combining blastulation timing and MC along with morphologic grading and euploid status may offer a new direction in embryo selection.
Appendix
Available only for authorised users
Literature
1.
go back to reference Veleva Z, Karinen P, Tomas C, Tapanainen JS, Martikainen H. Elective single embryo transfer with cryopreservation improves the outcome and diminishes the costs of IVF/ICSI. Hum Reprod. 2009;24:1632–9.CrossRefPubMed Veleva Z, Karinen P, Tomas C, Tapanainen JS, Martikainen H. Elective single embryo transfer with cryopreservation improves the outcome and diminishes the costs of IVF/ICSI. Hum Reprod. 2009;24:1632–9.CrossRefPubMed
2.
go back to reference Papanikolaou EG, Kolibianakis EM, Tournaye H, Venetis CA, Fatemi H, Tarlatzis B, et al. Live birth rates after transfer of equal number of blastocysts or cleavage-stage embryos in IVF. A systematic review and meta-analysis. Hum Reprod. 2008;23:91–9.CrossRefPubMed Papanikolaou EG, Kolibianakis EM, Tournaye H, Venetis CA, Fatemi H, Tarlatzis B, et al. Live birth rates after transfer of equal number of blastocysts or cleavage-stage embryos in IVF. A systematic review and meta-analysis. Hum Reprod. 2008;23:91–9.CrossRefPubMed
3.
go back to reference Gardner DK, Schoolcraft WB, Wagley L, Schlenker T, Stevens J, Hesla J. A prospective randomized trial of blastocyst culture and transfer in in vitro fertilization. Hum Reprod. 1998;13:3434–40.CrossRefPubMed Gardner DK, Schoolcraft WB, Wagley L, Schlenker T, Stevens J, Hesla J. A prospective randomized trial of blastocyst culture and transfer in in vitro fertilization. Hum Reprod. 1998;13:3434–40.CrossRefPubMed
4.
go back to reference Scott RT Jr, Upham KM, Forman EJ, Hong KH, Scott KL, Taylor D, et al. Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates: a randomized controlled trial. Fertil Steril. 2013;100:697–703.CrossRefPubMed Scott RT Jr, Upham KM, Forman EJ, Hong KH, Scott KL, Taylor D, et al. Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates: a randomized controlled trial. Fertil Steril. 2013;100:697–703.CrossRefPubMed
5.
go back to reference Fragouli E, Spath K, Alfarawati S, Kaper F, Craig A, Michel CE, et al. Altered levels of mitochondrial DNA are associated with female age, aneuploidy, and provide an independent measure of embryonic implantation potential. PLoS Genet. 2015;11:e1005241.CrossRefPubMedPubMedCentral Fragouli E, Spath K, Alfarawati S, Kaper F, Craig A, Michel CE, et al. Altered levels of mitochondrial DNA are associated with female age, aneuploidy, and provide an independent measure of embryonic implantation potential. PLoS Genet. 2015;11:e1005241.CrossRefPubMedPubMedCentral
6.
go back to reference Diez-Juan A, Rubio C, Marin C, Martinez S, Al-Asmar N, Riboldi M, et al. Mitochondrial DNA content as a viability score in human euploid embryos: less is better. Fertil Steril. 2015;104:534–41.CrossRefPubMed Diez-Juan A, Rubio C, Marin C, Martinez S, Al-Asmar N, Riboldi M, et al. Mitochondrial DNA content as a viability score in human euploid embryos: less is better. Fertil Steril. 2015;104:534–41.CrossRefPubMed
7.
go back to reference Cree LM, Samuels DC, de Sousa Lopes SC, Rajasimha HK, Wonnapinij P, Mann JR, et al. A reduction of mitochondrial DNA molecules during embryogenesis explains the rapid segregation of genotypes. Nat Genet. 2008;40:249–54.CrossRefPubMed Cree LM, Samuels DC, de Sousa Lopes SC, Rajasimha HK, Wonnapinij P, Mann JR, et al. A reduction of mitochondrial DNA molecules during embryogenesis explains the rapid segregation of genotypes. Nat Genet. 2008;40:249–54.CrossRefPubMed
8.
go back to reference Hashimoto S, Morimoto N, Yamanaka M, Matsumoto H, Yamochi T, Goto H, et al. Quantitative and qualitative changes of mitochondrial in human preimplantation embryos. J Assist Reprod Genet. 2017;34:573–80.CrossRefPubMedPubMedCentral Hashimoto S, Morimoto N, Yamanaka M, Matsumoto H, Yamochi T, Goto H, et al. Quantitative and qualitative changes of mitochondrial in human preimplantation embryos. J Assist Reprod Genet. 2017;34:573–80.CrossRefPubMedPubMedCentral
9.
go back to reference Shamsi MB, Govindaraj P, Chawla, Malhotra N, Singh N, Mittal S, et al. Mitochondrial DNA variations in ova and blastocyst: implications in assisted reproduction. Mitochondrion. 2013;13:96–105.CrossRefPubMed Shamsi MB, Govindaraj P, Chawla, Malhotra N, Singh N, Mittal S, et al. Mitochondrial DNA variations in ova and blastocyst: implications in assisted reproduction. Mitochondrion. 2013;13:96–105.CrossRefPubMed
10.
go back to reference Dumollard R, Duchen M, Carroll J. The role of mitochondrial function in theoocyte and embryo. Curr Top Dev Biol. 2007;77:21–49.CrossRefPubMed Dumollard R, Duchen M, Carroll J. The role of mitochondrial function in theoocyte and embryo. Curr Top Dev Biol. 2007;77:21–49.CrossRefPubMed
11.
go back to reference Wong CC, Loweke KE, Bossert NL, Behr B, De Jonge CJ, Baer TM, et al. Non-invasive imaging of human embryos before embryonic genome activation predicts development to the blastocyst stage. Nat Biotechnol. 2010;28:1115–21.CrossRefPubMed Wong CC, Loweke KE, Bossert NL, Behr B, De Jonge CJ, Baer TM, et al. Non-invasive imaging of human embryos before embryonic genome activation predicts development to the blastocyst stage. Nat Biotechnol. 2010;28:1115–21.CrossRefPubMed
12.
go back to reference Liu Y, Chapple V, Feenan K, Roberts PR, Matson P. Time-lapse deselection model for human day 3 in vitro fertilization embryos: the combination of qualitative and quantitative measures of embryo growth. Fertil Steril. 2016;105:656–62.CrossRefPubMed Liu Y, Chapple V, Feenan K, Roberts PR, Matson P. Time-lapse deselection model for human day 3 in vitro fertilization embryos: the combination of qualitative and quantitative measures of embryo growth. Fertil Steril. 2016;105:656–62.CrossRefPubMed
13.
go back to reference Kirkegaard K, Agerholm IE, Ingerslev HJ. Time-lapse monitoring as a tool for clinical embryo assessment. Hum Reprod. 2012;27:1277–85.CrossRefPubMed Kirkegaard K, Agerholm IE, Ingerslev HJ. Time-lapse monitoring as a tool for clinical embryo assessment. Hum Reprod. 2012;27:1277–85.CrossRefPubMed
14.
go back to reference Chawla M, Fakih M, Shunnar A, Bayram A, Hellani A, Perumal V, et al. Morphokinetic analysis of cleavage stage embryos and its relationship to aneuploidy in a retrospective time-lapse imaging study. J Assist Reprod Genet. 2015;32:69–75.CrossRefPubMed Chawla M, Fakih M, Shunnar A, Bayram A, Hellani A, Perumal V, et al. Morphokinetic analysis of cleavage stage embryos and its relationship to aneuploidy in a retrospective time-lapse imaging study. J Assist Reprod Genet. 2015;32:69–75.CrossRefPubMed
15.
go back to reference Racowsky C, Vernon M, Mayer J, Ball GD, Behr B, Pomeroy KO, et al. Standardization of grading embryo morphology. J Assist Reprod Genet. 2010;27:437–9.CrossRefPubMedPubMedCentral Racowsky C, Vernon M, Mayer J, Ball GD, Behr B, Pomeroy KO, et al. Standardization of grading embryo morphology. J Assist Reprod Genet. 2010;27:437–9.CrossRefPubMedPubMedCentral
16.
go back to reference Alpha Scientists in Reproductive Medicine and ESHRE Special Interest Group of Embryology. The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting. Hum Reprod. 2011;6:1270–83.CrossRef Alpha Scientists in Reproductive Medicine and ESHRE Special Interest Group of Embryology. The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting. Hum Reprod. 2011;6:1270–83.CrossRef
17.
go back to reference Campbell A, Fishel S, Bowman N, Duffy S, Sedler M, Lindermann Hickman CF. Modelling a risk classification of aneuploidy in human embryos using non-invasive morphokinetics. Repro Biomed Online. 2013;26:477–85.CrossRef Campbell A, Fishel S, Bowman N, Duffy S, Sedler M, Lindermann Hickman CF. Modelling a risk classification of aneuploidy in human embryos using non-invasive morphokinetics. Repro Biomed Online. 2013;26:477–85.CrossRef
20.
go back to reference Chavez SL, Loewke KE, Han J, Moussavi F, Colls P, Munne S, et al. Dynamic blastomere behavior reflects human embryo ploidy by the four-cell stage. Nat Commun. 2012;3:1251.CrossRefPubMedPubMedCentral Chavez SL, Loewke KE, Han J, Moussavi F, Colls P, Munne S, et al. Dynamic blastomere behavior reflects human embryo ploidy by the four-cell stage. Nat Commun. 2012;3:1251.CrossRefPubMedPubMedCentral
21.
go back to reference Basile N, del Carmen NM, Bronet F, Florensa M, Riquerios M, Rodrigo L, et al. Increasing the probability of selecting chromosomally normal embryos by time-lapse morphokinetics analysis. Fertil Steril. 2014;101:699–704.CrossRefPubMed Basile N, del Carmen NM, Bronet F, Florensa M, Riquerios M, Rodrigo L, et al. Increasing the probability of selecting chromosomally normal embryos by time-lapse morphokinetics analysis. Fertil Steril. 2014;101:699–704.CrossRefPubMed
22.
go back to reference Steuerwald N, Barritt JA, Adler R, Malter H, Schimmel T, Cohen J, et al. Quantification of mtDNA in single oocytes, polar bodies and subcellular components by real-time rapid cycle fluorescence monitored PCR. Zygote. 2000;8:209–15.CrossRefPubMed Steuerwald N, Barritt JA, Adler R, Malter H, Schimmel T, Cohen J, et al. Quantification of mtDNA in single oocytes, polar bodies and subcellular components by real-time rapid cycle fluorescence monitored PCR. Zygote. 2000;8:209–15.CrossRefPubMed
23.
go back to reference Barritt JA, Kokot M, Cohen J, Steuerwald N, Brenner CA. Quantification of human ooplasmic mitochondria. Reprod BioMed Online. 2002;4:243–7.CrossRefPubMed Barritt JA, Kokot M, Cohen J, Steuerwald N, Brenner CA. Quantification of human ooplasmic mitochondria. Reprod BioMed Online. 2002;4:243–7.CrossRefPubMed
24.
go back to reference St. John JC, Facucho-Oliveira J, Jiang Y, Kelly R, Salah R. Mitochondrial DNA transmission, replication and inheritance: a journey from the gamete through the embryo and into offspring and embryonic stem cells. Hum Reprod Update. 2010;16:488–509.CrossRefPubMed St. John JC, Facucho-Oliveira J, Jiang Y, Kelly R, Salah R. Mitochondrial DNA transmission, replication and inheritance: a journey from the gamete through the embryo and into offspring and embryonic stem cells. Hum Reprod Update. 2010;16:488–509.CrossRefPubMed
25.
go back to reference Thundathil J, Filion F, Smith LC. Molecular control of mitochondrial function in preimplantation mouse embryos. Mol Reprod Dev. 2005;71:405–13.CrossRefPubMed Thundathil J, Filion F, Smith LC. Molecular control of mitochondrial function in preimplantation mouse embryos. Mol Reprod Dev. 2005;71:405–13.CrossRefPubMed
26.
go back to reference Monnot S, Samuels DC, Hesters L, Frydman N, Gigarel N, Burlet P, et al. Mutation dependence of the mitochondrial DNA copy number in the first stages of human embryogenesis. Hum Mol Genet. 2013;22:1867–72.CrossRefPubMed Monnot S, Samuels DC, Hesters L, Frydman N, Gigarel N, Burlet P, et al. Mutation dependence of the mitochondrial DNA copy number in the first stages of human embryogenesis. Hum Mol Genet. 2013;22:1867–72.CrossRefPubMed
27.
go back to reference Victor AR, Brake AJ, Tyndall JC, Griffin DK, Zouves CG, Barnes FL, et al. Accurate quantitation of mitochondrial DNA reveals uniform levels in human blastocysts irrespective of ploidy, age, or implantation potential. Fertil Steril. 2016;107:34–42.CrossRefPubMed Victor AR, Brake AJ, Tyndall JC, Griffin DK, Zouves CG, Barnes FL, et al. Accurate quantitation of mitochondrial DNA reveals uniform levels in human blastocysts irrespective of ploidy, age, or implantation potential. Fertil Steril. 2016;107:34–42.CrossRefPubMed
28.
go back to reference Treff NR, Zhan Y, Tao X, Olcha M, Han M, Rajchel J, et al. Levels of trophectoderm mitochondrial DNA do not predict the reproductive potential of sibling embryos. Hum Reprod. 2017;32:954–62.PubMedPubMedCentral Treff NR, Zhan Y, Tao X, Olcha M, Han M, Rajchel J, et al. Levels of trophectoderm mitochondrial DNA do not predict the reproductive potential of sibling embryos. Hum Reprod. 2017;32:954–62.PubMedPubMedCentral
29.
go back to reference Thompson SM, Onwubalili, Brown K, Jindal SK, McGovern PG. Blastocyst expansion score and trophectoderm morphology strongly predict successful clinical pregnancy and live birth following elective single embryo blastocyst transfer (eSET): a national study. Journ Assist Reprod Genet. 2013;30:1577–81.CrossRef Thompson SM, Onwubalili, Brown K, Jindal SK, McGovern PG. Blastocyst expansion score and trophectoderm morphology strongly predict successful clinical pregnancy and live birth following elective single embryo blastocyst transfer (eSET): a national study. Journ Assist Reprod Genet. 2013;30:1577–81.CrossRef
30.
go back to reference Lin DP, Huang CC, HM W, Cheng TC, Chen CI, Lee MS. Comparison of mitochondrial DNA contents in human embryos with good or poor morphology at the 8-cell stage. Fertil Steril. 2004;81:73–9.CrossRefPubMed Lin DP, Huang CC, HM W, Cheng TC, Chen CI, Lee MS. Comparison of mitochondrial DNA contents in human embryos with good or poor morphology at the 8-cell stage. Fertil Steril. 2004;81:73–9.CrossRefPubMed
Metadata
Title
Blastulation timing is associated with differential mitochondrial content in euploid embryos
Authors
Jacqueline R. Ho
Nabil Arrach
Katherine Rhodes-Long
Wael Salem
Lynda K. McGinnis
Karine Chung
Kristin A. Bendikson
Richard J. Paulson
Ali Ahmady
Publication date
01-04-2018
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 4/2018
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-018-1113-9

Other articles of this Issue 4/2018

Journal of Assisted Reproduction and Genetics 4/2018 Go to the issue