Cell-to-Cell Adhesion and Electrical Parameters of Murine Embryo Membranes After Cryopreservation by Vitrification

Q4 Medicine Problems of Cryobiology and Cryomedicine Pub Date : 2022-12-30 DOI:10.15407/cryo32.04.256

Y. Smolyaninova, V. Shigimaga, Lyudmila Popivnenko, I. Kovalenko

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Abstract

In this work, we have studied the impact of various stages of cryopreservation by vitrification in ethylene glycolsucrose medium on plasma membranes adhesive properties of 8-cell murine embryos and their specific electrical conductivity. Embryos were divided into the following experimental groups: the control, group 1 (incubation in vitrification medium) and group 2 (complete cycle of cryopreservation). The embryo exposure to vitrification medium did not affect their ability to cell-to-cell adhesion. After a complete cycle of cryopreservation, no embryo adhesion was observed. Electrical conductivity of embryo membranes was determined using the pulsed conductometry. After incubation in vitrification medium, their resistance to electric pulse was shown to decrease, that was manifested in the phenomenon of irreversible electric breakdown. The average values of electrical conductivity varied within the following ranges: ((12.1 ± 1.5)...(55.5 ± 2.6)), ((28.7 ± 5.7)...(44.± 8.9)), ((31.0 ± 9.3)...(87.9 ± 26.1)) μS/cm in the control, groups 1 and 2, respectively. These findings may be explained by appearance of first structural disorders in lipid bilayer of embryo membranes even at the stage of their incubation in vitrification medium. The lack of adhesive properties of blastomeres after vitrification-warming testified to a damage to the membrane adhesion proteins.

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小鼠胚胎膜玻璃化冷冻保存后的细胞间粘附和电参数

在这项工作中，我们研究了在乙烯-乙醇蔗糖培养基中玻璃化冷冻保存的不同阶段对8细胞小鼠胚胎质膜粘附性能及其比电导率的影响。胚胎分为以下实验组：对照组、第1组（玻璃化培养基中孵育）和第2组（冷冻保存完整周期）。胚胎暴露于玻璃化培养基不会影响其细胞间粘附的能力。经过一个完整的冷冻保存周期后，没有观察到胚胎粘附。用脉冲电导法测定胚胎膜的电导率。在玻璃化培养基中培养后，它们对电脉冲的电阻降低，表现为不可逆的电击穿现象。对照组1和2的电导率平均值分别在以下范围内变化：（12.1±1.5）…（55.5±2.6））、（28.7±5.7）…（44.±8.9）、（31.0±9.3）…（87.9±26.1））μS/cm。这些发现可以通过胚胎膜脂质双层中首次出现结构紊乱来解释，即使在胚胎膜在玻璃化培养基中孵育的阶段也是如此。玻璃化加温后卵裂球缺乏粘附特性，证明膜粘附蛋白受损。

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来源期刊

Problems of Cryobiology and Cryomedicine Medicine-Medicine (miscellaneous)

CiteScore

0.60

自引率

0.00%

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期刊介绍： The Journal publishes the reviews and original papers on cryobiological and cryomedical research, in particular the elucidation of mechanisms of injuries occurring in biological objects and caused by the influence of low and ultra low temperatures; natural resistance of biologicals to cold and their recovery post effect; the development of effective methods of cryoprotection and technology of storage of biological resources under hypothermic and ultra low temperatures, application of hypothermia, cryotherapy and cryopreserved biologicals for treating various pathologies; cell and tissue based therapies and other issues of low-temperature biology and medicine, as well as development of devices and equipment for low temperature biology and medicine. The journal covers all topics related to low temperature biology, medicine and engineering. These include but are not limited to: low temperature storage of biologicals (human, animal or plant cells, tissues, and organs), including preparation for storage, thawing/warming, cell and tissue culturing etc. response of biologicals to low temperature; cold adaptation of animals and plants; utilisation of low temperature in medicine; experimental and clinical transplantation, cell and tissue based therapies; developing of cryobiological and cryomedical devices; organisation and functioning of low temperature banks etc.