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

IF 1 4区物理与天体物理 Q4 PHYSICS, APPLIED High Temperature Pub Date : 2024-03-20 DOI:10.1134/s0018151x23040144

D. S. Sitnikov, D. E. Mukhdina, D. S. Korshunova, Yu. Yu. Silaeva, M. A. Filatov, I. V. Ilina

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Abstract

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%.

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胚胎透明带激光显微切割对冷冻保存方案下胚胎透明带变薄动态的影响

摘要作者研究了在囊胚期进行激光辅助孵化导致小鼠胚胎透明带变薄的动态变化。小鼠胚胎先前经过冻融循环（冷冻胚胎），被选为模型胚胎。ZP 显微手术使用飞秒激光脉冲（辐射波长 512 nm，脉冲持续时间 100 fs，强度 2.5 TW/cm2）。在囊胚期（~E3.5，即胚胎发育的 3.5 天）和孵化期（~E5）进行显微手术前测量了 ZP 的厚度。结果发现，对照组胚胎（未接受激光照射的冷冻胚胎）的 ZP 变薄（从 6.6（E3.5）微米到 4.9 微米（E5）），而实验组胚胎在接受激光辅助孵化程序后的 ZP 变薄（从 7.1（E3.5）微米到 6.4 微米（E5））。在第一种和第二种情况下，ZP 厚度的变化都具有显著的统计学意义。这些结果与未进行冷冻保存的 "新鲜 "胚胎的数据进行了比较。与新鲜胚胎相比，冷冻胚胎的 ZP 硬化效果并不明显。与对照组胚胎相比，冷冻胚胎使用激光辅助孵化程序可将成功孵化的概率从 38.5%提高到 52.5%。

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来源期刊

High Temperature 物理-物理：应用

CiteScore

1.50

自引率

40.00%

发文量

审稿时长

4-8 weeks

期刊介绍： High Temperature is an international peer reviewed journal that publishes original papers and reviews written by theoretical and experimental researchers. The journal deals with properties and processes in low-temperature plasma; thermophysical properties of substances including pure materials, mixtures and alloys; the properties in the vicinity of the critical point, equations of state; phase equilibrium; heat and mass transfer phenomena, in particular, by forced and free convections; processes of boiling and condensation, radiation, and complex heat transfer; experimental methods and apparatuses; high-temperature facilities for power engineering applications, etc. The journal reflects the current trends in thermophysical research. It presents the results of present-day experimental and theoretical studies in the processes of complex heat transfer, thermal, gas dynamic processes, and processes of heat and mass transfer, as well as the latest advances in the theoretical description of the properties of high-temperature media.