Influence of Laser Microdisection of the Zona Pellucida of Embryos on the Its Thinning Dynamics under Cryopreservation Protocols

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The authors studied the dynamics of thinning of the zona pellucida (ZP) in mouse embryo as a result of laser-assisted hatching performed at the blastocyst stage. Mouse embryos previously subjected to a freeze–thaw cycle (cryo-embryos) were selected as model embryos. For ZP microsurgery, femtosecond laser pulses were used (radiation wavelength 512 nm, pulse duration 100 fs, intensity 2.5 TW/cm2). The thickness of the ZP was measured before microsurgery at the blastocyst stage (~E3.5, i.e., 3.5 days of embryonic development) and at the hatching stage (~E5). It was found that the ZP of control group embryos (cryo-embryos not subjected to laser exposure) thinned more strongly (from 6.6 (E3.5) to 4.9 µm (E5)) compared to experimental embryos after the laser-assisted hatching procedure (from 7.1 ( E3.5) to 6.4 µm (E5)). In both the first and second cases, changes in ZP thickness were statistically significant. The results were compared with data for “fresh” embryos not subjected to cryopreservation. There was no pronounced effect of ZP hardening in cryo-embryos compared to fresh embryos. The use of the laser-assisted hatching procedure for cryo-embryos made it possible to increase the probability of successful hatching compared to embryos in the control group from 38.5 to 52.5%.

作者简介

D. Sitnikov

Joint Institute for High Temperatures, Russian Academy of Sciences

Email: Sitnik.ds@gmail.com
Moscow, Russia

D. Mukhdina

Joint Institute for High Temperatures, Russian Academy of Sciences

Email: Sitnik.ds@gmail.com
Moscow, Russia

D. Korshunova

Joint Institute for High Temperatures, Russian Academy of Sciences; Institute of Gene Biology, Russian Academy of Sciences

Email: Sitnik.ds@gmail.com
Moscow, Russia; Moscow, Russia

Yu. Silaeva

Joint Institute for High Temperatures, Russian Academy of Sciences; Institute of Gene Biology, Russian Academy of Sciences

Email: Sitnik.ds@gmail.com
Moscow, Russia; Moscow, Russia

M. Filatov

Joint Institute for High Temperatures, Russian Academy of Sciences; Institute of Gene Biology, Russian Academy of Sciences

Email: Sitnik.ds@gmail.com
Moscow, Russia; Moscow, Russia

I. Ilina

Joint Institute for High Temperatures, Russian Academy of Sciences

编辑信件的主要联系方式.
Email: Sitnik.ds@gmail.com
Moscow, Russia

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版权所有 © Д.С. Ситников, Д.Е. Мухдина, Д.С. Коршунова, Ю.Ю. Силаева, М.А. Филатов, И.В. Ильина, 2023