家畜卵母细胞玻璃化冷冻保护剂应用进展与优化策略

doi:10.16431/j.cnki.1671-7236.2026.02.001

摘要/Abstract

摘要：

卵母细胞玻璃化冷冻保存技术是保存优良畜禽种质资源和珍稀濒危物种的重要手段，也是推动品种改良和胚胎产业化生产的重要技术支撑。冷冻保护剂是玻璃化冷冻的关键媒介，通过降低溶液冰点、增加溶液黏度、抑制冰晶形成和重结晶、缓解渗透损伤等机制发挥保护作用。然而，冷冻保护剂的细胞毒性是玻璃化冷冻中亟待解决的核心问题之一。开发低毒、高效的新型冷冻保护剂已成为提高家畜卵母细胞玻璃化冷冻效率的重要研究方向。文章系统分析了传统冷冻保护剂对卵母细胞的毒性损伤机制，重点综述了小分子渗透调节剂、脂质调节剂、抗氧化剂及细胞骨架稳定剂等新型冷冻保护剂的应用进展，旨在为优化家畜卵母细胞玻璃化冷冻体系提供理论依据和技术参考。

关键词: 卵母细胞; 玻璃化冷冻; 冷冻损伤; 冷冻保护剂

Abstract:

Oocyte cryopreservation is a vital technology for preserving germplasm resources of genetically superior livestock and endangered species， and also serving as a critical tool for breed improvement and commercial embryo production. As essential agents in vitrification， cryoprotectant agents protect cells by lowering the freezing point， increasing viscosity， preventing ice crystallization， and mitigating osmotic damage. However， the cytotoxicity of cryoprotectant agents remains one of the critical challenges to be addressed in vitrification technology. The development of novel cryoprotectant agents with low toxicity and high efficacy has become a key research direction for improving the efficiency of livestock oocyte vitrification. This review systematically analyzes the cytotoxic mechanisms of conventional cryoprotectant agents in oocytes preservation， with particular emphasis on recent advances in novel cryoprotective agents， including small-molecule permeable cryoprotectants， lipid modulators， antioxidants， and cytoskeleton stabilizers. It will provide both theoretical basis and technical references for optimizing the vitrification systems of livestock oocytes.

Key words: oocytes; vitrification; cryoinjury; cryoprotectant agents

中图分类号:

S814

李伉, 陈思颍, 孙雅雯, 冷璇, 王栋, 庞云渭. 家畜卵母细胞玻璃化冷冻保护剂应用进展与优化策略[J]. 中国畜牧兽医, 2026, 53(2): 521-531.

LI Kang, CHEN Siying, SUN Yawen, LENG Xuan, WANG Dong, PANG Yunwei. Application Advance and Optimization Strategies in Cryoprotectant Agents for Vitrification of Livestock Oocytes[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(2): 521-531.

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小分子渗透调节剂 Small-molecule permeable cryoprotectants	物种 Species	冷冻阶段 Freezing stage	处理方式Treatments	评价指标 Evaluation indicators	冷冻对照组 Frozen control group	冷冻处理组 Frozen treatment group	参考文献 References
甘氨酸 Glycine	猪	GV	IVF	囊胚率	2.5%	11.0%	Tang等^［24］
	单峰骆驼	GV	IVF	囊胚率	12.0%	18.5%	Yaqout等^［27］
	绵羊	GV	IVF	囊胚率	6.2%	10.4%	Ahmadi等^［28］
脯氨酸 Proline	猪	GV	PA	囊胚率	5.4%	13.5%	孙雅雯^［33］
谷氨酰胺 Glutamine	牛	GV	—	细胞核成熟率	17.4%	32.8%	Yamada等^［21］

脂质调节剂 Lipid modulators	物种 Species	冷冻阶段 Freezing stage	处理方式Treatments	评价指标 Evaluation indicators	冷冻对照组 Frozen control group	冷冻处理组 Frozen treatment group	参考文献 References
毛喉素 Forskolin	猪	MⅡ	PA	存活率	51.7%	65.3%	Fu等^［38］
毛喉素 Forskolin	牛	GV	IVF	囊胚率	10.8%	31.7%	Ezoe等^［47］
反式10，顺式共轭亚油酸 Trans-10， cis-12 conjugated linoleic acid	牛	MⅡ	IVF	存活率	71.7%	80.4%	Matos等^［43］
L-肉碱 L-carnitine	牛	MⅡ	IVF	囊胚率	20.2%	34.4%	Chankitisakul 等^［48］
β-烟酰胺 β-nicotinamide mononucleotide	牛	MⅡ	IVF	囊胚率	15.4%	33.75%	Xu等^［36］
草虫素 Cordycepin	牛	MⅡ	IVF	囊胚率	13.2%	23.6%	Xu等^［36］
小檗碱 Berberine	牛	MⅡ	IVF	囊胚率	12.5%	25.0%	Xu等^［36］

抗氧化剂 Antioxidants	物种 Species	冷冻阶段 Freezing stage	处理方式Treatments	评价指标 Evaluation indicators	冷冻对照组 Frozen control group	冷冻处理组 Frozen treatment group	参考文献 References
白藜芦醇 Resveratrol	牛	MⅡ	IVF	卵裂率	26.7%	40.5%	Gutierrez-Castillo等^［59］
	牛	GV	IVF	囊胚率	15.8%	33.3%	Zhang等^［60］
	猪	GV	PA	存活率	60.6%	77.3%	Ito等^［71］
	猪	GV	PA	囊胚率	21.2%	32.6%	Santos等^［72］
褪黑素 Melatonin	猪	GV	IVF	囊胚率	2.5%	14.2%	Tang等^［24］
虾青素 Astaxanthin	猪	GV	PA	囊胚率	14.1%	21.3%	Xiang等^［67］
辅酶Q10 Coenzyme Q10	牛	GV	—	存活率	57.9%	77.2%	Ruiz-Conca 等^［56］
槲皮素 Quercetin	绵羊	GV	IVF	囊胚率	7.0%	25.0%	Davoodian 等^［57］

细胞骨架稳定剂 Cytoskeleton stabilizers	物种 Species	冷冻阶段 Freezing stage	处理方式 Treatments	评价指标 Evaluation indicators	冷冻对照组 Frozen control group	冷冻处理组 Frozen treatment group	参考文献 References
紫杉醇 Paclitaxel	牛	MⅡ	IVF	卵裂率	34.0%	41.9%	Morató等^［78］
	猪	MⅡ	—	存活率	54.6%	60.67%	Fu等^［80］
	猪	MⅡ	PA	卵裂率	5.6%	24.3%	Shi等^［81］
	猪	MⅡ	PA	囊胚率	8.3%	18.6%	Ogawa等^［85］
多西他赛 Docetaxel	牛	MⅡ	IVF	囊胚率	27.3%	39.3%	Pitchayapipatkul 等^［74］
细胞松弛素B Cytochalasin B	猪	MⅡ	—	存活率	35.1%	44.2%	Hwang等^［84］