Effect of laser optoperforation of the zona pellucida on mouse embryo development in vitro

Zakharchenko, E. O.; Zalessky, A. D.; Osychenko, A. A.; Krivokharchenko, Alexander; Shakhbazyan, A. K.; Ryabova, A. V.; Надточенко, В. А.

doi:10.1134/s0006297915060127

Cited by 3 publications

(2 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This indicates the safety of laser device on assisted hatching. Zakharchenko et al reported that zona pellucida perforation by laser device at the blas-tocyst stage had a negative effect and that hatching did not occur after perforation of zona pellucida [18]. Therefore, we could assume that the safety or detrimental effect of laser device on blastocyst is dependent on the method of laser application.…”

Section: Discussionmentioning

confidence: 94%

Which is the safer method for trophectoderm biopsy in mouse blastocyst, mechanical or laser?

2020

Clinical and Experimental Obstetrics & Gynecology

View full text Add to dashboard Cite

Introduction: This study was conducted to compare hatching rates after assisted hatching, re-expansion rates after trophectoderm biopsy, and survival rates after cryopreservation using different methods of assisted hatching and biopsy in mouse embryo. Materials and Methods: Five-week-old female mice (C57BL/CBA) were superovulated, and two-cell embryos were collected. All embryos were cultured to blastocyst stage. For assisted hatching and separating trophectoderm from blastocyst, laser device and hand-made pipette were used respectively. Hatching rates after assisted hatching, re-expansion rates after trophectoderm biopsy, and survival rates after cryopreservation were calculated. Results: Hatching rate was 92% in mechanically assisted hatching group and 90% in laser group, respectively. After mechanically assisted hatching, re-expansion rate was 91.3% and survival rate was 87% in biopsy by pipette and laser group, respectively. In laser hatching group, re-expansion rate was 88.9% with biopsy by pipette and survival rate was 84.4% with biopsy by laser. Conclusion: Throughout the study, mechanical technique and laser technique showed no differences in the safety profiles in trophectoderm biopsy procedure.

show abstract

Section: Discussionmentioning

confidence: 94%

Which is the safer method for trophectoderm biopsy in mouse blastocyst, mechanical or laser?

2020

Clinical and Experimental Obstetrics & Gynecology

View full text Add to dashboard Cite

show abstract

“…The results of ZP drilling at blastocyst stage in locations close to either the TE or ICM have recently been reported [42][43][44][52][53][54][55][56] and are summarized in Table 3. As a rule, an infrared diode laser (λ = 1.48 μm) is used to create a hole.…”

Section: Discussionmentioning

confidence: 99%

Controlled hatching at the prescribed site using femtosecond laser for zona pellucida drilling at the early blastocyst stage

Ilina

Khramova

Ivanova

et al. 2020

J Assist Reprod Genet

View full text Add to dashboard Cite

Purpose To study whether the application of femtosecond laser pulses for zona pellucida (ZP) drilling of blastocysts at the embryonic or abembryonic poles can promote hatching to start immediately through the hole formed and ensure high hatching rates and embryo viability. Methods Mouse blastocyst (E3.5) ZP were microdissected with femtosecond laser pulses (514-nm wavelength, 280-fs pulse duration, 2.5-kHz repetition rate) close to the trophoblast or inner cell mass (ICM). The sizes of the holes formed were in the range of 4.5-8.5 μm. Additional longitudinal incisions (5-7-μm long) on either side of the hole were created to determine whether hatching had started at the correct position. Embryos post-laser-assisted ZP drilling and intact embryos were cultured under standard conditions for 2 days; embryo quality was assessed twice daily. The hatching rates and in vitro and in vivo implantation rates (only for embryos with ZP dissected close to the ICM) were estimated. Results Femtosecond laser-assisted ZP drilling at the early blastocyst stage facilitated embryo hatching to start at the artificial opening with probability approaching 100%. Despite the artificial opening's small size, no embryo trapping during hatching was observed. Both experimental groups had higher hatching rates than the control groups (93.3-94.7% vs. 83.3-85.7%, respectively). The in vitro implantation rate was comparable with that of the control group (92.3% vs. 95.4%). No statistically significant differences were obtained in the in vivo implantation rates between the experimental and control groups. Conclusions Blastocyst-stage femtosecond laser microsurgery of ZP is fast and delicate and enables the hatching process to be initiated in a controlled manner through a relatively small opening, with no embryo trapping.

show abstract

Assisted Hatching

Shinde¹

2022

Advances in Assisted Reproduction Technologies

View full text Add to dashboard Cite

The selected Spermatozoa when it reaches the ovulated Cumulus Oocyte Complex after ovulation, dispersion of the granulosa cells and corona radiata cells occur. The Spermatozoa then must cross the Zona Pellucida (ZP), fuse with the oolemma, and then subsequently fertilize the oocyte. Embryologist Karl Ernst von Baer coined the term ‘Zona Pellucida’ from Greek work Zone which means belt or girdle and Latin work Pellucida which means transparent or shining. This extracellular matrix is about 13-15 um thick and surrounds all the mammalian eggs and pre-implantation embryos. Zona Pellucida structure is made up of carbohydrates, specific proteins, glycoproteins, hyaluronic acid, heparin, collagen, and fibrous proteins. Human Zona Pellucida contains 4 glycosylated proteins namely ZP1, ZP2, ZP3, and ZP4. ZP plays an important role in helping oocytes to transport essential nutrients and helps in avoiding polyspermy by hardening after fertilization. The embryos must break open the protective ZP layer to the implant, the process is called hatching. It is said that in Assisted reproductive treatment (ART) factors such as the non-availability of enzymes from the endometrium which helps in hatching, extended culture, vitrification may lead to failure in the hatching of embryos from ZP. It was postulated that micromanipulation of ZP to create an opening will help the embryos to hatch and thus implant and will lead to an increase in Implantation rates (IR). This process was later called Assisted Hatching (AH). Various methods were discovered for Assisted hatching such as mechanical ZP AH, zona digestion using enzymes, and laser-Assisted hatching. This chapter will focus on the advantages and disadvantages of each method of AH and their applications in ART along with the impact of AH on clinical outcomes. The use of any method of AH should be chosen carefully to avoid damage to the embryo which will defy the whole purpose of application of AH. In any case, laser-assisted hatching is widely used for Pre- Implantation Genetic Testing (PGT) of the embryos as it is very safe if applied properly, convenient, easy to use, and faster compared to other methods of AH. Each laboratory should identify the correct time and stage at which application of AH is considered based on whether it is helping to improve clinical rates or not.

show abstract

Effect of laser optoperforation of the zona pellucida on mouse embryo development in vitro

Cited by 3 publications

References 16 publications

Which is the safer method for trophectoderm biopsy in mouse blastocyst, mechanical or laser?

Which is the safer method for trophectoderm biopsy in mouse blastocyst, mechanical or laser?

Controlled hatching at the prescribed site using femtosecond laser for zona pellucida drilling at the early blastocyst stage

Assisted Hatching

Contact Info

Product

Resources

About