取样后动物胚胎的冷冻保存技术研究进展

doi:10.11843/j.issn.0366-6964.2025.10.004

Abstract

Abstract:

Post-sampling embryo cryopreservation technology is a critical component of preimplantation genetic testing (PGT), holding significant application value in embryo sex selection, genetic screening, genomic selection, and assisted reproduction. However, embryonic sampling procedures may lead to zona pellucida damage, reduced cell count, and epigenetic abnormalities, thereby limiting cryopreservation efficiency. Recent advancements, including optimized sampling methods (e.g., polar body biopsy, blastomere biopsy, and trophectoderm biopsy), improved culture medium formulations (e.g., supplementation with L-arginine and glutathione), and refined cryopreservation protocols (e.g., vitrification techniques), have markedly enhanced the cryopreservation efficiency of post-sampling embryos. Furthermore, non-invasive techniques (e.g., analysis of cell-free DNA in embryo culture medium) and epigenetic modifications (e.g., DNA methylation regulation) offer novel approaches to minimize embryonic damage. This review summarizes the mechanisms of cryopreservation-induced damage in post-sampling embryos and protective strategies, with a focus on cutting-edge advancements in zona pellucida repair, epigenetic regulation, and cryopreservation efficiency. By evaluating the strengths and limitations of existing technologies, this review aims to provide theoretical foundations and technical references for developing safe and efficient cryopreservation protocols, thereby promoting their application and adoption in livestock breeding and assisted reproductive technologies.

Key words: embryo sampling, PGT, cryopreservation, freezing efficiency

CLC Number:

S814

DONG Jianhua, YANG Baigao, ZHANG Xiaomeng, FENG Xiaoyi, SONG Haoran, LIU Yang, WANG Zizhuo, WANG Yanbo, LI Chongyang, LÜ Lihua, ZHAO Xueming. Research Progress on Cryopreservation Technology of Animal Embryos after Sampling[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 4796-4806.

Figures/Tables 4

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Table 1

References 78

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物种 Species	推荐活检阶段 Recommendation for biopsy stage	取样细胞类型 Sample cell type	技术特点 Technical characteristics	参考文献 Reference
牛Cattle	囊胚期	滋养外胚层(TE)	激光辅助透明带打孔，取5~8个细胞；玻璃化冷冻技术成熟。	[40]
小鼠Mouse	卵裂期(8细胞)	卵裂球	显微操作针机械取样，单细胞活检；冻存耐受性差，需改良冷冻液。	[41]
人类Human	囊胚期	滋养外胚层(TE)	激光脉冲精准切割，取5~10个细胞；玻璃化冷冻后存活率可达90%以上。	[42]
猪Pig	囊胚期	滋养外胚层(TE)	透明带厚且弹性差，需高强度激光或酶解法辅助打孔；冷冻保存技术落后。	[43]
绵羊Sheep	囊胚期	滋养外胚层(TE)	激光辅助取样，活检后短期培养(5~8 h)可提升冷冻存活率。	[44]
山羊Goat	囊胚期	滋养外胚层(TE)	透明带较薄，激光打孔后机械吸取；冷冻耐受性中等。	[45]
兔Rabbit	卵裂期(8细胞)	卵裂球	胚胎体积较大(120~150 μm)，操作相对容易；冻存耐受性中等。	[46]
猴Monkey	囊胚期	滋养外胚层(TE)	非侵入性cfDNA分析为主，避免伦理争议；激光取样技术成熟。	[47]
斑马鱼Zebrafish	卵裂期(16-32细胞)	卵裂球	胚胎透明，易于观察；显微注射技术成熟。	[48]

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Research Progress on Cryopreservation Technology of Animal Embryos after Sampling

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