Skip to main content
SpringerLink
Log in

Laser-assisted hatching of cleavage-stage embryos impairs developmental potential and increases DNA damage in blastocysts

  • Original Article
  • Published:
Lasers in Medical Science Aims and scope Submit manuscript

Abstract

This study aims to investigate the influence of two- (day 2) and six-to-eight-cell-stage (day 3) laser-assisted hatchings on the developmental potential and genetic integrity of the embryos. In this prospective experimental study, two- and six-to-eight-cell-stage mouse embryos were subjected to laser hatching using 1,480 nm diode laser, and then assessed for the developmental potential and DNA integrity in blastocysts. Similarly, four-cell-stage human embryos from 20 patients were also subjected to laser hatching, and then assessed for the developmental competence. Laser-assisted hatching in mouse embryos significantly enhanced the blastocyst hatching potential on day 4.5 (P < 0.0001). However, a significant decline in blastocyst total cell number (TCN) was observed in six-to-eight-cell-stage laser-hatched embryos (P < 0.001). Conversely, no significant difference in TCN was observed between laser-hatched and unhatched human four-cell-stage embryos after 24 h. Attempt to understand the genetic integrity in laser-hatched mouse blastocysts revealed significantly higher labeling index when hatching was done at two- (P < 0.01) and six-to-eight-cell stage (P < 0.05). DNA damage induced by the laser manipulation may affect implantation and postimplantation developmental potential of the embryos. However, further studies are required to elucidate the impact of laser-induced DNA damage on the reproductive outcome.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Mansour RT, Rhodes CA, Aboulghar MA, Serour GI, Kamal A (2000) Transfer of zona-free embryos improves outcome in poor prognosis patients: a prospective randomized controlled study. Hum Reprod 15:1061–1064

    Article  CAS  PubMed  Google Scholar 

  2. Cohen J, Alikani M, Trowbridge J, Rosenwaks Z (1992) Implantation enhancement by selective assisted hatching using zona drilling of human embryos with poor prognosis. Hum Reprod 7:685–691

    CAS  PubMed  Google Scholar 

  3. Stein A, Rufas O, Amit S, Avrech O, Pinkas H, Ovadia J, Ovadia J, Fisch B (1995) Assisted hatching by partial zona dissection of human pre-embryos in patients with recurrent implantation failure after in vitro fertilization. Fertil Steril 63:838–841

    CAS  PubMed  Google Scholar 

  4. Bider D, Livshits A, Yonish M, Yemini Z, Mashiach S, Dor J (1997) Assisted hatching by zona drilling of human embryos in women of advanced age. Hum Reprod 12:317–320

    Article  CAS  PubMed  Google Scholar 

  5. Lanzendorf SE, Nehchiri F, Mayer JF, Oehninger S, Muasher SJ (1998) A prospective, randomized, double-blind study for the evaluation of assisted hatching in patients with advanced maternal age. Hum Reprod 13:409–413

    Article  CAS  PubMed  Google Scholar 

  6. Practice Committee of Society for Assisted Reproductive Technology; Practice Committee of American Society for Reproductive Medicine (2008) The role of assisted hatching in in vitro fertilization: a review of the literature. A Committee opinion. Fertil Steril 90:S196–S198

    Google Scholar 

  7. Cohen J (1993) Assisted hatching: indications and techniques. Acta Eur Fertil 24:215–219

    CAS  PubMed  Google Scholar 

  8. Cohen J, Alikani M, Reing AM, Ferrara TA, Trowbridge J, Tucker M (1992) Selective assisted hatching of human embryos. Ann Acad Med Singap 21:565–570

    CAS  PubMed  Google Scholar 

  9. Antinori S, Panci C, Selman HA, Caffa B, Dani G, Versaci C (1996) Zona thinning with the use of laser: a new approach to assisted hatching in humans. Hum Reprod 11:590–594

    Article  CAS  PubMed  Google Scholar 

  10. Balaban B, Urman B, Alatas C, Mercan R, Mumcu A, Isiklar A (2002) A comparison of four different techniques of assisted hatching. Hum Reprod 17:1239–1243

    Article  PubMed  Google Scholar 

  11. Hsieh YY, Huang CC, Cheng TC, Chang CC, Tsai HD, Lee MS (2002) Laser-assisted hatching of embryos is better than the chemical method for enhancing the pregnancy rate in women with advanced age. Fertil Steril 78:179–182

    Article  PubMed  Google Scholar 

  12. Makrakis E, Angeli I, Agapitou K, Pappas K, Dafereras A, Pantos K (2006) Laser versus mechanical assisted hatching: a prospective study of clinical outcomes. Fertil Steril 86:1596–1600

    Article  PubMed  Google Scholar 

  13. Rink K, Delacrétaz G, Salathé RP, Senn A, Nocera D, Germond M, De Grandi P, Fakan S (1996) Non-contact microdrilling of mouse zona pellucida with an objective-delivered 1.48-microns diode laser. Lasers Surg Med 18:52–62

    Article  CAS  PubMed  Google Scholar 

  14. Levin I, Almog B, Shwartz T, Gold V, Ben-Yosef D, Shaubi M, Amit A, Malcov M (2012) Effects of laser polar-body biopsy on embryo quality. Fertil Steril 97:1085–1088

    Article  PubMed  Google Scholar 

  15. Wong BC, Boyd CA, Lanzendorf SE (2003) Randomized controlled study of human zona pellucida dissection using the zona infrared laser optical system: evaluation of blastomere damage, embryo development, and subsequent hatching. Fertil Steril 80:1249–1254

    Article  PubMed  Google Scholar 

  16. Montag M, van der Ven H (1999) Laser-assisted hatching in assisted reproduction. Croat Med J 40:398–403

    CAS  PubMed  Google Scholar 

  17. Han TS, Sagoskin AW, Graham JR, Tucker MJ, Liebermann J (2003) Laser-assisted human embryo biopsy on the third day of development for preimplantation genetic diagnosis: two successful case reports. Fertil Steril 80:453–455

    Article  PubMed  Google Scholar 

  18. Hartshorn C, Anshelevich A, Wangh LJ (2005) Laser zona drilling does not induce hsp70i transcription in blastomeres of eight-cell mouse embryos. Fertil Steril 84:1547–1550

    Article  CAS  PubMed  Google Scholar 

  19. Hurst BS, Tucker KE, Awoniyi A, Schlaff WD (1998) Assisted hatching does not enhance IVF success in good-prognosis patients. J Assist Reprod Genet 15:62–64

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  20. Douglas-Hamilton DH, Conia J (2001) Thermal effects in laser-assisted pre-embryo zona drilling. J Biomed Opt 6:205–213

    Article  CAS  PubMed  Google Scholar 

  21. Ealy AD, Howell JL, Monterroso VH, Aréchiga CF, Hansen PJ (1995) Developmental changes in sensitivity of bovine embryos to heat shock and use of antioxidants as thermoprotectants. J Anim Sci 73:1401–1407

    CAS  PubMed  Google Scholar 

  22. Adiga SK, Toyoshima M, Shiraishi K, Shimura T, Takeda J, Taga M, Nagai H, Kumar P, Niwa O (2007) p21 provides stage specific DNA damage control to preimplantation embryos. Oncogene 26:6141–6149

    Article  CAS  PubMed  Google Scholar 

  23. Adiga SK, Toyoshima M, Shimura T, Takeda J, Uematsu N, Niwa O (2007) Delayed and stage specific phosphorylation of H2AX during preimplantation development of gamma-irradiated mouse embryos. Reproduction 133:415–422

    Article  CAS  PubMed  Google Scholar 

  24. Newport J, Kirschner M (1982) A major developmental transition in early Xenopus embryos: II. Control of the onset of transcription. Cell 30:687–696

    Article  CAS  PubMed  Google Scholar 

  25. Jeong YJ, Choi HW, Shin HS, Cui XS, Kim NH, Gerton GL, Jun JH (2005) Optimization of real time RT-PCR methods for the analysis of gene expression in mouse eggs and preimplantation embryos. Mol Reprod Dev 71:284–289

    Article  CAS  PubMed  Google Scholar 

  26. Malter HE, Schimmel T, Cohen J (2001) Zona dissection by infrared laser: developmental consequences in the mouse, technical considerations, and controlled clinical trial. Reprod BioMed Online 3:117–123

    Article  PubMed  Google Scholar 

  27. Flach G, Johnson MH, Braude PR, Taylor RA, Bolton VN (1982) The transition from maternal to embryonic control in the 2-cell mouse embryo. EMBO J 1:681–686

    CAS  PubMed Central  PubMed  Google Scholar 

  28. Montag M, Koll B, Holmes P, Van der Ven H (2000) Significance of the number of embryonic cells and the state of the zona pellucida for hatching of mouse blastocysts in vitro versus in vivo. Biol Reprod 62:1738–1744

    Article  CAS  PubMed  Google Scholar 

  29. Heyer BS, MacAuley A, Behrendtsen O, Werb Z (2000) Hypersensitivity to DNA damage leads to increased apoptosis during early mouse development. Genes Dev 14:2072–2084

    CAS  PubMed Central  PubMed  Google Scholar 

  30. Weil M, Jacobson MD, Coles HS, Davies TJ, Gardner RL, Raff KD, Raff MC (1996) Constitutive expression of the machinery for programmed cell death. J Cell Biol 133:1053–1059

    Article  CAS  PubMed  Google Scholar 

  31. Chandrakanthan V, Li A, Chami O, O’Neill C (2006) Effects of in vitro fertilization and embryo culture on TRP53 and Bax expression in B6 mouse embryos. Reprod Biol Endocrinol 4:61

    Article  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by Indian Council of Medical Research (ICMR) grant no. 2010-00020.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Division of Clinical Embryology, Department of Obstetrics and Gynecology, Kasturba Medical College, Manipal University, Manipal, 576 104, India

    Sachin D. Honguntikar, Shubhashree Uppangala, Sujith R. Salian, Guruprasad Kalthur & Satish K. Adiga

  2. Department of Obstetrics and Gynecology, Kasturba Medical College, Manipal University, Manipal, 576 104, India

    Pratap Kumar

Authors
  1. Sachin D. Honguntikar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shubhashree Uppangala
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sujith R. Salian
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Guruprasad Kalthur
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Pratap Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Satish K. Adiga
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Satish K. Adiga.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Honguntikar, S.D., Uppangala, S., Salian, S.R. et al. Laser-assisted hatching of cleavage-stage embryos impairs developmental potential and increases DNA damage in blastocysts. Lasers Med Sci 30, 95–101 (2015). https://doi.org/10.1007/s10103-014-1625-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10103-014-1625-1

Keywords

Search

Navigation