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

Abstract

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

CLC Number:

S814

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.

Figures/Tables 4

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References 85

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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］

Application Advance and Optimization Strategies in Cryoprotectant Agents for Vitrification of Livestock Oocytes

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[1]	LI Zhuo, WANG Pei, GUO Jianxiong, ZHANG Xiong, ZHAO Hongfang, JIAO Deling, JIA Baoyu, QING Yubo, CHENG Wenmin, KUI Hua, WEI Hongjiang. Effects of Porcine Oocyte Morphology and Quality on the Development of Somatic Cell Nuclear Transfer Embryo [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(12): 5785-5796.
[2]	MO Xianhong, LI Junjie, GUO Cheng, WEN Zhaoyu, ZOU Yuzhu, ZHAO Wenbo, XU Zhenjun. Research Progress on Vitrification Injury of Oocytes in Livestock [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(6): 2524-2532.
[3]	LUO Anfeng, HUA Zaidong, YANG Caixia, CHEN Fan. Effect of Nuclear Membrane Pore Subcomplex Nup98/Rae1 on Meiotic Maturation of Mouse Oocytes in vitro [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(5): 1998-2006.
[4]	LI Youwei, CHENG Yazhuo, SHANG Jiyong, ZHANG Tinglong, SUN Mingju. Research Progress on in vitro Maturation of Small Follicle-derived Oocytes in Mammalian [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(3): 1171-1182.
[5]	LIU Keke, XUE Mengqi, TANG Feitai, DU Qiansheng, WANG Xinyu, DONG Yinyu, WANG Xiaoli. Effect of AFP Ⅲ on Mitochondria of Vitrified Mouse Oocytes [J]. China Animal Husbandry & Veterinary Medicine, 2023, 50(8): 3180-3188.
[6]	XIANG Decai, LI Shuiying, ZHANG Bin, ZHANG Yan, LIANG Jiachong, LYU Chunrong, HONG Qionghua, QUAN Guobo, WU Guoquan. The Study on the Role of Fetal Bovine Serum Supplementation During Postwarming Culture on Vitrified Porcine Parthenogenetic Blastocysts [J]. China Animal Husbandry & Veterinary Medicine, 2023, 50(2): 606-615.
[7]	MO Xianhong, YUE Kaiping, SUN Liyao, LI Bing, ZHAO Bing, GUO Cheng, XU Zhenjun. Effect of 2-APB on Development Capacity of Vitrified Bovine Mature Oocytes [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(7): 2669-2676.
[8]	JI Wenhui, WANG Yuling, HE Honghong, FU Wei, LAN Daoliang. Effects of Vitamin A on the Maturation and Subsequent Development of Yak Oocytes in vitro [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(12): 4707-4714.
[9]	JIAO Anhui, ZHANG Xiaomeng, ZHAO Yuhan, WANG Yu, GAO Qingshan. Effects of Limonin on in vitro Maturation of Mouse Oocytes and Embryo Development of in vitro Fertilization [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(10): 3891-3900.
[10]	WANG Yu, ZHAO Yuhan, JIAO Anhui, WANG Yuqi, GAO Qingshan. Effects of Acetochlor on in vitro Maturation of Mouse Oocytes [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(10): 3909-3917.
[11]	LI Xiaoxia, XIAO Hongwei, CAO Pinghua, XU Zhiqian, ZHANG Fang, ZHANG Zhiyang, JING Penghua, YU Xueli, LI Yinghua. Effects of Trehalose on Vitrification of Bovine Immature Oocytes [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(1): 224-231.
[12]	YAO Ying, CHEN Yan, XIONG Xianrong, MIPAM Tserang-donko, CHAI Zhixin, JI Wenhui, LAN Daoliang. Study on the Expression and Subcellular Localization of G Protein-coupled Receptor 50 During in vitro Maturation Process of Yak Oocytes [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(9): 3387-3393.
[13]	XUE Mengqi, ZHOU Yue, LIU Keke, WANG Xinyu, DONG Yinyu, TANG Xiaochuan, WANG Xiaoli. Effect of Vitrification on MⅡ Oocytes and Their DNA of Porcine [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(7): 2475-2483.
[14]	ZHU Tianqi, LI Jia, LIU Guoshi, ZHANG Lu. The Roles of Calcium Oscillations in the Fertilization and Activation of Oocytes [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(4): 1343-1350.
[15]	YAO Yujun, LI Guangdong, WU Hao, MA Wenkui, YANG Hai, GUAN Shengyu, LYU Dongying, FU Yao, ZHU Tianqi, JI Pengyun, TAN Xinxing, ZHAO Wanmin, LIAN Zhengxing, ZHANG Lu, LIU Guoshi. Effects of Resveratrol and Melatonin on Transfection of Sheep Fetal Fibroblasts and in vitro Maturation of Oocytes [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(12): 4497-4507.