College of Animal Science and Technology, Beijing University of Agriculture, Beijing, 102206, China.
College of Economics and Management, Beijing University of Agriculture, Beijing, 102206, China.
Reprod Biol. 2022 Mar;22(1):100612. doi: 10.1016/j.repbio.2022.100612. Epub 2022 Feb 2.
The cryopreservation of mammalian embryos is an important technology in embryo engineering. The discovery and application of the embryo's own high freezing resistance factors are the main methods to improve the utilization of mammalian embryos in cryopreservation. Cathepsin L gene expression in the frozen and thawed dormant embryos displayed a significant difference from those normal hatched ones. The aim of the present study was to dig out the potential role of Cathepsin L in anti-freezing capacity of murine blastocysts by investigating the location and expression of Cathepsin L in frozen and thawed both activated and dormant hatching blastocysts. Different concentrations of Cathepsin L recombinant protein and E-64d were then respectively added into the embryo cryoprotectant and pre-cryo culture medium. Our results found that down-regulation of Cathepsin L improves the freezing resistance of murine normal hatching embryos by reducing apoptosis. Cathepsin L inhibitors can be used to improve the efficiency of cryopreservation and recovery of blastocysts in vitro. Our study provides a theoretical basis for the further development and application of Cathepsin L.
哺乳动物胚胎的冷冻保存是胚胎工程中的一项重要技术。发现和应用胚胎自身的高抗冻性因子是提高哺乳动物胚胎在冷冻保存中利用率的主要方法。冷冻和解冻休眠胚胎中组织蛋白酶 L 的表达与正常孵化胚胎有显著差异。本研究旨在通过研究冷冻和解冻激活和休眠孵化囊胚中组织蛋白酶 L 的位置和表达，挖掘组织蛋白酶 L 在囊胚抗冻能力中的潜在作用。然后将不同浓度的组织蛋白酶 L 重组蛋白和 E-64d 分别添加到胚胎冷冻保护剂和预冷冻培养中。研究结果发现，下调组织蛋白酶 L 可通过减少凋亡来提高正常孵化胚胎的抗冻性。组织蛋白酶 L 抑制剂可用于提高体外冷冻保存和囊胚恢复的效率。本研究为进一步开发和应用组织蛋白酶 L 提供了理论基础。
