马卵母细胞玻璃化冷冻保存研究进展

doi:10.11843/j.issn.0366-6964.2025.09.002

Abstract

Abstract:

Oocyte cryopreservation is of significant importance for human assisted reproductive technology and livestock production.This technology aids in the conservation of genetic resources from superior breeds and the maintenance of biodiversity.However, due to their small surface area-to-volume ratio and low membrane permeability, oocytes are highly susceptible to damage during freezing.This includes mechanical injury caused by ice crystal formation, oxidative stress, and the toxicity of cryoprotectants, all of which severely reduce their developmental competence after thawing. Compared to other animals, the high lipid content within equine oocytes reduces their cryotolerance, making cryopreservation considerably more challenging. Although offspring can be obtained from vitrified immature oocytes, the developmental competence of vitrified-thawed oocytes remains to be enhanced compared to their fresh oocytes.This article primarily reviews the types of damage caused by vitrification to equine oocytes and the factors influencing the cryopreservation efficiency of equine oocytes.It aims to provide a reference for further establishing and optimizing vitrification protocols for equine oocyte cryopreservation.

Key words: equine, oocytes, vitrification, cryodamage, cryoprotective agent

CLC Number:

S821.35

YANG Mingying, WANG Na, LIU Yuanyi, LI Xinyu, Bayanamar , SHI Yujie, MANG Lai, DU Ming. Research Progress on Vitrification Cryopreservation of Equine Oocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4143-4155.

Figures/Tables 6

Table 1

Table 2

Fig. 1

Table 3

Table 4

Table 5

References 75

1	CASILLAS F , BETANCOURT M . The porcine experimental model for oocyte cryopreservation by vitrification[J]. AJBSR, 2020, 9 (2): 165- 169. doi: 10.34297/AJBSR.2020.09.001377
2	BOGLIOLO L , LEDDA S , INNOCENZI P , et al. Raman microspectroscopy as a non-invasive tool to assess the vitrification-induced changes of ovine oocyte zona pellucida[J]. Cryobiology, 2012, 64 (3): 267- 272. doi: 10.1016/j.cryobiol.2012.02.010
3	MARTEIL G , METCHAT A , DOLLET S , et al. Vitrification of human oocytes before or after Rescue-IVM does not impair maturation kinetics but induces meiotic spindle alterations[J]. Reprod Sci, 2024, 31 (9): 2625- 2636. doi: 10.1007/s43032-024-01596-7
4	SUN H , GUO Y , YU R , et al. Ru360 protects against vitrification-induced oocyte meiotic defects by restoring mitochondrial function[J]. Theriogenology, 2023, 204, 40- 49. doi: 10.1016/j.theriogenology.2023.04.004
5	ZHANG C , ZHAO M , XUE Y , et al. Effects of vitrification on mitochondrial ultrastructure and membrane potential and its distribution in mouse oocytes[J]. Cryo Lett, 2024, 45 (5): 301- 308. doi: 10.54680/fr24510110212
6	PEREIRA B C , ORTIZ I , DORADO J , et al. Evaluation of DNA damage of mare granulosa cells before and after cryopreservation using a chromatin dispersion test[J]. J Equine Vet Sci, 2019, 72, 28- 30. doi: 10.1016/j.jevs.2018.10.019
7	ORTIZ-ESCRIBANO N , BOGADO PASCOTTINI O , WOELDERS H , et al. An improved vitrification protocol for equine immature oocytes, resulting in a first live foal[J]. Equine Vet J, 2018, 50 (3): 391- 397. doi: 10.1111/evj.12747
8	WHITTINGHAM D G . Fertilization in vitro and development to term of unfertilized mouse oocytes previously stored at -196 degrees C[J]. J Reprod Fertil, 1977, 49 (1): 89- 94. doi: 10.1530/jrf.0.0490089
9	FUKU E , KOJIMA T , SHIOYA Y , et al. In vitro fertilization and development of frozen-thawed bovine oocytes[J]. Cryobiology, 1992, 29 (4): 485- 492. doi: 10.1016/0011-2240(92)90051-3
10	LE GAL F . In vitro maturation and fertilization of goat oocytes frozen at the germinal vesicle stage[J]. Theriogenology, 1996, 45 (6): 1177- 1185. doi: 10.1016/0093-691X(96)00073-8
11	DIDION B A , POMP D , MARTIN M J , et al. Observations on the cooling and cryopreservation of pig oocytes at the germinal vesicle stage[J]. J Anim Sci, 1990, 68 (9): 2803- 2810. doi: 10.2527/1990.6892803x
12	RALL W F , FAHY G M . Ice-free cryopreservation of mouse embryos at -196℃ by vitrification[J]. Nature, 1985, 313 (6003): 573- 575. doi: 10.1038/313573a0
13	NAKAGATA N . High survival rate of unfertilized mouse oocytes after vitrification[J]. J Reprod Fertil, 1989, 87 (2): 479- 483. doi: 10.1530/jrf.0.0870479
14	HOCHI S , FUJIMOTO T , CHOI Y H , et al. Cryopreservation of equine oocytes by 2-step freezing[J]. Theriogenology, 1994, 42 (7): 1085- 1094. doi: 10.1016/0093-691X(94)90856-7
15	MACLELLAN L J , CARNEVALE E M , COUTINHO DA SILVA M A , et al. Pregnancies from vitrified equine oocytes collected from super-stimulated and non-stimulated mares[J]. Theriogenology, 2002, 58 (5): 911- 999. doi: 10.1016/S0093-691X(02)00920-2
16	CANESIN H S , BROM-DE-LUNA , J G , CHOI Y H , et al. Blastocyst development after intracytoplasmic sperm injection of equine oocytes vitrified at the germinal-vesicle stage[J]. Cryobiology, 2021, 75, 52- 59.
17	CLÉRICO G , TAMINELLI G , VERONESI J C , et al. Mitochondrial function, blastocyst development and live foals born after ICSI of immature vitrified/warmed equine oocytes matured with or without melatonin[J]. Theriogenology, 2021, 160, 40- 49. doi: 10.1016/j.theriogenology.2020.10.036
18	CHEN C . Pregnancy after human oocyte cryo-preservation[J]. Lancet, 1986, 1 (8486): 884- 886.
19	AL-HASANI S , KIRSCH J , DIEDRICH K , et al. Successful embryo transfer of cryopreserved and in-vitro fertilized rabbit oocytes[J]. Hum Reprod, 1989, 4 (1): 77- 79. doi: 10.1093/oxfordjournals.humrep.a136849
20	CANESIN H S , ORTIZ I , ROCHA FILHO A N , et al. Effect of warming method on embryo quality in a simplified equine embryo vitrification system[J]. Theriogenology, 2020, 151, 151- 158. doi: 10.1016/j.theriogenology.2020.03.012
21	AI-HASANI S , KIRSCH J , DIEDRICH K , et al. Successcul embryo transfer of cryopreserved and in-vitro fertilized rabbit oocytes[J]. Hum Reprod, 1989, 4 (1): 77- 79. doi: 10.1093/oxfordjournals.humrep.a136849
22	KONO T , KWON O Y , NAKAHARA T . Development of enucleated mouse oocytes reconstituted with embryonic nuclei[J]. J Reprod Fertil, 1991, 93, 154- 172.
23	RUBINSKY B , ARAV A , DEVRIES A L . The cryoprotective effect of antifreeze glycopeptides from Antarctic fishes[J]. Cryobiology, 1992, 29 (1): 69- 79. doi: 10.1016/0011-2240(92)90006-N
24	HAMANO S , KOIKEDA A , KUWAYAMA M , et al. Full-term development of in vitro-matured, vitrified and fertilized bovine oocytes[J]. Theriogenology, 1992, 38 (6): 1085- 1090. doi: 10.1016/0093-691X(92)90122-8
25	NAGY Z P , CHANG C C , SHAPIRO D B , et al. Clinical evaluation of the efficiency of an oocytedonation program using egg cryo-banking[J]. Fert Ster, 2009, 92 (2): 520- 526. doi: 10.1016/j.fertnstert.2008.06.005
26	DATTENA M , PTAK G , LOI P , et al. Survival and viability of vitrified in vitro and in vivo produced ovine blastocysts[J]. Theriogenology, 2000, 53 (8): 1511- 1519. doi: 10.1016/S0093-691X(00)00293-4
27	THARASANIT T , COLENBRANDER B , STOUT T A . Effect of maturation stage at cryopreservation on post-thaw cytoskeleton quality and fertilizability of equine oocytes[J]. Mol Reprod Dev, 2006, 73 (5): 627- 637. doi: 10.1002/mrd.20432
28	THARASANIT T , COLLEONI S , LAZZARI G , et al. Effect of cumulus morphology and maturation stage on the cryopreservability of equine oocytes[J]. Reproduction, 2006, 132 (5): 759- 769. doi: 10.1530/rep.1.01156
29	MACLELLAN L J , STOKES J E , PREIS K A , et al. Vitrification, warming, ICSI and transfer of equine oocytes matured invivo[J]. Anim Reprod Sci, 2010, 121, 5260- 5261.
30	NOWAK A , KOCHAN J , PAPIS K , et al. Studies on survival of horse oocytes after Rapid-i method vitrification[J]. J Equ Vet Sci, 2014, 34 (5): 675- 679. doi: 10.1016/j.jevs.2013.12.013
31	DE COSTER T , VELEZ D A , VAN SOOM A , et al. Cryopreservation of equine oocytes: looking into the crystal ball[J]. Reprod Fertil Dev, 2020, 32 (5): 453- 467. doi: 10.1071/RD19229
32	CANESIN H S , BROM-DE-LUNA J G , CHOI Y H , et al. Vitrification of germinal-vesicle stage equine oocytes: Effect of cryoprotectant exposure time on in-vitro embryo production[J]. Cryobiology, 2018, 81, 185- 191. doi: 10.1016/j.cryobiol.2018.01.001
33	DUCHEYNE K D , RIZZO M , DAELS P F , et al. Vitrifying immature equine oocytes impairs their ability to correctly align the chromosomes on the MⅡ spindle[J]. Reprod Fertil Dev, 2019, 31 (8): 1330- 1338. doi: 10.1071/RD18276
34	PEREIRA B C , ORTIZ I , DORADO J M , et a . Effect of permeable cryoprotectant-free vitrification on DNA fragmentation of equine oocyte-cumulus cells[J]. Reprod Domest Anim, 2019, 54 (Suppl 3): 53- 56.
35	LÓPEZ A , DUCOLOMB Y , CASAS E , et al. Effects of porcine immature oocyte vitrification on actin microfilament distribution and chromatin integrity during early embryo development in vitro[J]. Front Cell Dev Biol, 2021, 9, 636765. doi: 10.3389/fcell.2021.636765
36	MACLELLAN L J , ALBERTINI DF , STOKES J E , et al. Use of confocal microscopy and intracytoplasmic sperm injection (ICSI) to assess viability of equine oocytes from young and old mares after vitrification[J]. J Assist Reprod Genet, 2023, 40 (11): 2565- 2576. doi: 10.1007/s10815-023-02935-4
37	YURCHUK T , LIKSZO P , WITEK K , et al. New approach to the cryopreservation of GV oocytes and cumulus cells through the lens of preserving the intercellular gap junctions based on the bovine model[J]. Int J Mol Sc, 2024, 25 (11): 60- 74.
38	THARASANIT T , COLLEONI S , GALLI C , et al. Protective effects of the cumulus-corona radiata complex during vitrification of horse oocytes[J]. Reproduction, 2009, 137 (3): 391- 401. doi: 10.1530/REP-08-0333
39	GOOK D A , OSBORN S M , JOHNSTON W , et al. Cryopreservation of mouse and human oocytes using 1, 2-propanediol and the configuration of the meiotic spindle[J]. Hum Reprod, 1993, 8 (7): 1101- 1109. doi: 10.1093/oxfordjournals.humrep.a138201
40	VINCENT C , PICKERING S J , JOHNSON M H , et al. The hardening effect of dimethylsulphoxide on the mouse zona pellucida requires the presence of an oocyte and is associated with a reduction in the number of cortical granules present[J]. J Reprod Fertil, 1990, 89 (1): 253- 259. doi: 10.1530/jrf.0.0890253
41	LARMAN M G , SHEEHAN C B , GARDNER D K , et al. Calcium-free vitrification reduces cryoprotectant-induced zona pellucida hardening and increases fertilization rates in mouse oocytes[J]. Reproduction, 2006, 131 (1): 53- 61. doi: 10.1530/rep.1.00878
42	CHOI J K , YUE T , HUANG H , et al. The crucial role of zona pellucida in cryopreservation of oocytes by vitrification[J]. Cryobiology, 2015, 71 (2): 350- 355. doi: 10.1016/j.cryobiol.2015.08.012
43	DUMOLLARD R , DUCHEN M , CARROLL J . The role of mitochondrial function in the oocyte and embryo[J]. Curr Top Dev Biol, 2007, 77, 21- 49.
44	ZHANG C , ZHAO M , XUE Y , et al. Effects of vitrification on mitochondrial ultrastructure and membrane potential and its distribution in mouse oocytes[J]. Cryo Letters, 2024, 45 (5): 301- 308. doi: 10.54680/fr24510110212
45	MA Y , LONG C , LIU G , et al. WGBS combined with RNA-seq analysis revealed that Dnmt1 affects the methylation modification and gene expression changes during mouse oocyte vitrification[J]. Theriogenology, 2022, 177, 11- 21. doi: 10.1016/j.theriogenology.2021.09.032
46	CHEN H , ZHANG L , DENG T , et al. Effects of oocyte vitrification on epigenetic status in early bovine embryos[J]. Theriogenology, 2016, 86 (3): 868- 878. doi: 10.1016/j.theriogenology.2016.03.008
47	孙雅雯, 陈思颍, 李伉, 等. 猪卵母细胞玻璃化冷冻损伤的缓解策略[J]. 畜牧兽医学报, 2025, 56 (1): 36- 44. doi: 10.11843/j.issn.0366-6964.2025.01.004
	SUN Y W , CHEN S Y , LI K , et al. Strategies for alleviating cryoinjury of porcine vitrified-oocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56 (1): 36- 44. doi: 10.11843/j.issn.0366-6964.2025.01.004
48	贾宝瑜. 共轭亚油酸对猪卵母细胞体外成熟、胚胎培养和冷冻保存的作用机制[D]. 北京: 中国农业大学, 2014.
	JIA B Y. Effects of t10c12 CLA on oocyte in vitro maturation, embryo development and cryopreservation in pig[D]. Beijing: China Agricultural University, 2014. (in Chinese)
49	WALTER J , COLLEONI S , LAZZARI G , et al. Maturational competence of equine oocytes is associated with alterations in their 'cumulome'[J]. Mol Hum Reprod, 2024, 30 (9): 33.
50	TORNER H , ALM H , KANITZ W , et al. Effect of initial cumulus morphology on meiotic dynamic and status of mitochondria in horse oocytes during IVM[J]. Reprod Domest Anim, 2007, 42 (2): 176- 183. doi: 10.1111/j.1439-0531.2006.00749.x
51	ANGEL D , CANESIN H S , BROM-DE-LUNA J G , et al. Embryo development after vitrification of immature and in vitro-matured equine oocytes[J]. Cryobiology, 2020, 92, 251- 254. doi: 10.1016/j.cryobiol.2020.01.014
52	KUWAYAMA M . Highly efficient vitrification for cryopreservation of human oocytes and embryos: The Cryotop method[J]. Theriogenology, 2007, 67 (1): 73- 80. doi: 10.1016/j.theriogenology.2006.09.014
53	SANSINENA M , SANTOS M V , ZARITZKY N , et al. Numerical simulation of cooling rates in vitrification systems used for oocyte cryopreservation[J]. Cryobiology, 2011, 63 (1): 32- 37. doi: 10.1016/j.cryobiol.2011.04.006
54	陈思颍, 孙雅雯, 李伉, 等. 微流体技术在家畜体外胚胎生产中的应用进展[J]. 畜牧兽医学报, 2023, 54 (12): 4889- 4897. doi: 10.11843/j.issn.0366-6964.2023.12.001
	CHEN S Y , SUN Y W , LI K , et al. Application of microfluidic technologies in livestock in vitro embryo production[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54 (12): 4889- 4897. doi: 10.11843/j.issn.0366-6964.2023.12.001
55	LI Y , ZHANG J , HAN W , et al. Multifunctional laser-induced graphene-based microfluidic chip for high-performance oocyte cryopreservation with low concentration of cryoprotectants[J]. Adv Healthc Mater, 2024, 13 (23): e2400981.
56	LEI Z , XIE D , MBOGBA M K , et al. A microfluidic platform with cell-scale precise temperature control for simultaneous investigation of the osmotic responses of multiple oocytes[J]. Lab Chip, 2019, 19 (11): 1929- 1940. doi: 10.1039/C9LC00107G
57	ZHOU X L , GUO Y Y , YI X Y , et al. Experimental study of microfluidic chip for cryopreservation of oocytes[J]. Prog Biochem Biophys, 2018, 45 (7): 763- 771.
58	AGNIESZKA N , JOANNA K , WOJCIECH W , et al. In vitro maturation of equine oocytes followed by two vitrification protocols and subjected to either intracytoplasmic sperm injection (ICSI) or parthenogenic activation[J]. Theriogenology, 2021, 162, 42- 48. doi: 10.1016/j.theriogenology.2020.12.022
59	SOMFAI T , DANG-NGUYEN T Q , KIKUCHI K . Altered microfilament dynamics contribute to the formation of diploid metaphase spindles in porcine oocytes which fail to reach the metaphase-Ⅱ stage during in vitro maturation[J]. Anim Sci J, 2022, 93 (1): e13690. doi: 10.1111/asj.13690
60	TONE M , UKYO R , SAKAMOTO S H , et al. Effects of paclitaxel before vitrification on the nuclear maturation and development of immature porcine oocytes[J]. Cryo Lett, 2023, 44 (5): 307- 313. doi: 10.54680/fr23510110812
61	董胤余, 王勇杰, 刘可可, 等. 抗冷冻蛋白Ⅲ对玻璃化冷冻猪GV期卵母细胞及其DNA的保护作用[J]. 中国兽医科学, 2023, 53 (5): 658- 663.
	DONG Y Y , WANG Y J , LIU K K , et al. Protection of antifreeze protein Ⅲ on vitrified GV oocytes and its dna of porcine[J]. Chinese Veterinary Science, 2023, 53 (5): 658- 663.
62	CHEEPA F F , LIU H , ZHAO G . The Natural cryoprotectant honey for fertility cryopreservation[J]. Bioengineering (Basel), 2022, 9 (3): 88. doi: 10.3390/bioengineering9030088
63	ISHII M , KAMOSHITA M , KURIHARA Y , et al. Successful production of offspring derived from mouse zygotes vitrified with carboxylated ε-poly-L-lysine and polyvinyl alcohol without serum[J]. J Reprod Dev, 2023, 69 (1): 53- 55. doi: 10.1262/jrd.2022-121
64	SCHWARZ K R L , DE CASTRO F C , SCHEFER L , et al. The role of cGMP as a mediator of lipolysis in bovine oocytes and its effects on embryo development and cryopreservation[J]. PLoS One, 2018, 13 (6): e0191023.
65	ZHUAN Q , LI J , DU X , et al. Antioxidant procyanidin B2 protects oocytes against cryoinjuries via mitochondria regulated cortical tension[J]. J Anim Sci Biotechnol, 2022, 16, 13.
66	DU X , LI J , ZHUAN Q , et al. Artificially increasing cortical tension improves mouse oocytes development by attenuating meiotic defects during vitrification[J]. Front Cell Dev Biol, 2022, 10, 876259. doi: 10.3389/fcell.2022.876259
67	ZHANG P , YANG S , ZHANG H , et al. Vitrification of bovine germinal vesicle oocytes significantly decreased the methylation level of their in vitro derived MⅡ oocytes[J]. Reprod Fertil Dev, 2022, 34 (13): 889- 903. doi: 10.1071/RD22130
68	IWATA H . Resveratrol enhanced mitochondrial recovery from cryopreservation-induced damages in oocytes and embryos[J]. Reprod Med Biol, 2021, 20 (4): 419- 426. doi: 10.1002/rmb2.12401
69	BARROS V R P , MONTE A P O , SANTOS J M S , et al. Melatonin improves development, mitochondrial function and promotes the meiotic resumption of sheep oocytes from in vitro grown secondary follicles[J]. Theriogenology, 2020, 144, 67- 73. doi: 10.1016/j.theriogenology.2019.12.006
70	DUJÍČKOVÁ L , OLEXIKOVÁ L , MAKAREVICH A V , et al. Astaxanthin added during post-warm recovery mitigated oxidative stress in bovine vitrified oocytes and improved quality of resulting blastocysts[J]. Antioxidants (Basel), 2024, 13 (5): 556. doi: 10.3390/antiox13050556
71	XIANG D C , JIA B Y , FU X W , et al. Role of astaxanthin as an efficient antioxidant on the in vitro maturation and vitrification of porcine oocytes[J]. Theriogenology, 2021, 167, 13- 23. doi: 10.1016/j.theriogenology.2021.03.006
72	MORATÓ R , IZQUIERDO D , ALBARRACÍN J L , et al. Effects of pre-treating in vitro-matured bovine oocytes with the cytoskeleton stabilizing agent taxol prior to vitrification[J]. Mol Reprod Dev, 2008, 75 (1): 191- 201. doi: 10.1002/mrd.20725
73	李婉君, 徐皆欢, 何孟纤, 等. 细胞松弛素B改善冷冻引起的猪卵母细胞皮质颗粒迁移障碍[J]. 畜牧兽医学报, 2024, 55 (5): 1999- 2010. doi: 10.11843/j.issn.0366-6964.2024.05.018
	LI W J , XU J H , HE M Q , et al. Cytochalasin B alleviates the migration disorder of cortical particle caused by vitrification in porcine oocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55 (5): 1999- 2010. doi: 10.11843/j.issn.0366-6964.2024.05.018
74	MOAWAD A R , ZHU J , CHOI I , et al. Effect of Cytochalasin B pretreatment on developmental potential of ovine oocytes vitrified at the germinal vesicle stage[J]. Cryo Lett, 2013, 34 (6): 634- 644.
75	DU M , LI X Y , BAYINNAMULA , et al. Optimization of vitrification methods for equine oocytes[J]. Tissue Cell, 2024, 91, 102632. doi: 10.1016/j.tice.2024.102632

方法 Method	研究者 Investigator	年份 Year	物种 Species	成就 Event	参考文献 Reference
慢速冷冻 Slow freezing	Whittingham	1977	小鼠	首次保存卵母细胞获得后代	[8]
	Chen	1986	人	保存卵母细胞获得后代	[18]
	Al-Hasani等	1989	兔	保存卵母细胞获得后代	[19]
	Didion等	1990	猪	成功保存卵母细胞	[11]
	Fuku等	1992	牛	保存卵母细胞获得后代	[9]
	Hochi等	1994	羊	成功保存卵母细胞	[14]
	Canesin等	2020	马	成功保存卵母细胞	[20]
玻璃化冷冻 Vitrification freezing	Al-Hasani等	1989	兔	成功保存卵母细胞	[21]
	Kono等	1991	小鼠	首次保存卵母细胞获得后代	[22]
	Rubinsky等	1992	猪	成功保存卵母细胞	[23]
	Hamano等	1992	牛	保存卵母细胞获得后代	[24]
	Hochi等	1994	马	成功保存卵母细胞	[14]
	Nagy等	2009	人	成功保存卵母细胞	[25]
	Dattena等	2000	羊	保存卵母细胞获得后代	[26]

研究者 Investigator	年份 Year	分裂阶段 Splitting stage	冷冻组合 Frozen combination	发育能力 Developmental capacity	参考文献 Reference
Hochi等	1994	GV	10% EG+10.0 mol·L^-1 Sucrose	成熟率15.8%	[14]
Maclellan等	2002	MII	ED/EDFS，Three-step Sucrose	存活率73%	[15]
Tharasanit等	2006	GV	10% EG+0.3 mol·L^-1Sucrose	成熟率35%，卵裂率8%	[27]
Tharasanit等	2006	GV	10% EG+10% DMSO	卵裂率34%，囊胚率16%	[28]
Tharasanit等	2006	MII	20% EG+20% DMSO+0.3 mol·L^-1Sucrose	卵裂率27%，囊胚率4%	[28]
Maclellan等	2010	MII	Commercial Cryotop medium	卵裂率83%，囊胚率40%，妊娠率26%	[29]
Nowak等	2014	MII	EquiPro-VitKit (Minitube) commercial medium	存活率63%，卵裂率10%	[30]
Canesin等	2021	GV	FBS+PG+EG+T；30 s	成熟率42%，卵裂率80%，囊胚率10%	[16]
De Coster等	2020	GV	BS+PE 40%+0.3 mol·L^-1Galactose	成熟率41.2%，卵裂率65.5%，囊胚率9.4%	[31]
Ortiz-Escribano等	2018	GV	HS+15% EG+15% DMSO+0.5 mol·L^-1Sucrose	成熟率25.3%，卵裂率30%，囊胚率5%	[7]
Canesin等	2018	GV	EP/EPT	成熟率21%，卵裂率85%，囊胚率15%	[32]

研究者 Author	分裂阶段 Splitting stage	试验组 Experimental group	载体 Carrier	存活率/% Survival rate	成熟率/% Maturity rate	卵裂率/% Cleavage rate	囊胚率/% Blastocyst rate	妊娠率/% Pregnancy rate
Maclellan等^[15]	MII	对照组	Nylon	100	-	-	-	83
Maclellan等^[15]	MII	ED/EDFS	Nylon	73	-	-	-	12
Tharasanit等^[27]	GV	对照组	OPS	96	56	66	17	-
	GV	ED/EDS		63	54	34	1	-
	MII	ED/EDS		32	54	16	0	-
Maclellan等^[29]	MII	商业Cryotop培养基	Cryotop	72	86	83	40	26
Canesin等^[16]	GV	对照组	Stainless	-	73	93	19	-
Canesin等^[16]	GV	ED/EDS	Steel mesh	-	36	67	11	-
De Coster等^[31]	GV	对照组	MVD	-	58	76	20	-
De Coster等^[31]	GV	ED/EDS	MVD	-	48	30	0	-

作者 Author	分裂阶段 Splitting stage	载体 Carrier	平衡液VS1 Buffered solution	冷冻液VS2 Refrigerating fluid	解冻方案 Thawing solution	成熟率/% Maturity rate	卵裂率/% Cleavage rate	囊胚率/% Blastocyst rate	妊娠率/% Pregnancy rate
Canesin等^[32]	GV	不锈钢网	2%EG+2%PG+HM；40 s	17.5%EG+17.5%PG+0.3 mol·L^-1海藻糖；65 s	BM培养基一步解冻、标准逐步解冻	21	85	15	-
Agnieszka等^[58]	MII	Rapid-Ⅰ	EquiPro VitKit；10 min	EquiPro VitKit；30 s	EquiPro VitKit培养基一步解冻	-	10.2	10	-
Maclellan等^[36]	MII	Cryotop	7.5%EG+7.5%DMSO+HM；1 min	15%EG+15%DMSO+0.5 mol·L^-1蔗糖+HM；1min	标准逐步解冻	-	32	32	-
Ortiz-Escribano等^[7]	GV	Cryotop	10% EG+10%DMSO+HM；25 s	20% EG+20%DMSO+0.3 mol·L^-1蔗糖；15 s	标准逐步解冻	34	42	7	20
Angel等^[51]	GV	尼龙网	FBS/7.5%PG-7.5%EG；10 min	15%PG-15%EG；30 s	标准逐步解冻	42	80	10	-
Clérico等^[17]	GV	Cryotop	5% EG+5% DMSO；45 s	20% EG+20% DMSO+0.65 mol·L^-1；30 s	标准逐步解冻	54	46	9	33
Ducheyne等^[33]	GV	接种环	20% EG+20% DMSO+BS；25 s	10% EG+10% DMSO+0.5 mol·L^-1蔗糖；15 s	标准逐步解冻	33	-	-	-
Tharasanit等^[28]	GV	OPS	10% EG+10%DMSO+HM；30 s	20% EG+20% DMSO+0.5 mol·L^-1；5 s	0.3 mol·L^-1蔗糖一步解冻法	54	34	1	-
Tharasanit等^[28]	MII	OPS	10% EG+10% DMSO+HM；30 s	20% EG+20% DMSO+0.5 mol·L^-1；15 s	0.3 mol·L^-1蔗糖一步解冻法	-	16	-	-

类别 Category		物种 Species	浓度 Concentration	成果 Outcome	参考文献 Reference
抗氧化剂 Antioxidant	原花青素 B2(PB2)	水牛	10^-9 mol·L^-1	PB2改善水牛卵母细胞的发育能力和线粒体分布	[65]
	原花青素 B2(PB2)	小鼠	5 μg·mL^-1	PB2改善线粒体功能、提高囊胚率	[66]
	白藜芦醇(Res)	牛	2 μmol·L^-1	Res恢复DNA甲基化水平并显著改善玻璃化GV卵母细胞的成熟和发育能力	[67]
	白藜芦醇(Res)	小鼠	0.1 μmol·L^-1	Res修复玻璃化后胚胎的异常线粒体分布和线粒体功能障碍	[68]
	褪黑素(MT)	绵羊	1 μmol·L^-1	MT降低活性氧水平并促进胚胎抗氧化酶(如谷胱甘肽)的产生	[69]
	褪黑素(MT)	马	10 mmol·L^-1	MT降低线粒体相关的ROS和改善ICSI胚胎发育，卵裂率从45%提高至60%，囊胚率也从8%提升至14%	[20]
	虾青素(Ax)	牛	5.3 μmol·L^-1	Ax减少氧化应激改善囊胚质量	[70]
	虾青素(Ax)	猪	2.5 μmol·L^-1	Ax恢复玻璃化卵母细胞基因表达提高mRNA水平	[71]
细胞骨架稳定剂 Cytoskeleton stabilizer	紫杉醇(Taxol)	牛	1 μmol·L^-1	添加Taxol后线粒体分布正常率40%显著高于对照组22%	[72]
	紫杉醇(Taxol)	猪	10 mol·L^-1	添加Taxol后玻璃化线粒体分布正常率83.69%显著高于对照组48.5%	[56]
	细胞松弛素B(CB)	猪	7.5 μg·mL^-1	添加CB使卵母细胞卵裂率5.64%提高至20.91%，囊胚率由0.41%提高至5.00%	[73]
	细胞松弛素B(CB)	绵羊	7.5 μg·mL^-1	CB玻璃化组的受精率高于未添加CB组(57.0%vs40.7%)并提升胚胎质量	[74]

Research Progress on Vitrification Cryopreservation of Equine Oocytes

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 75

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	TANG Yu, ZHANG Ying, YANG Yifeng, XUE Hailong, LIU Lixiang, XU Baozeng. Mechanisms of Glycine Improving Vitrification Cryopreservation Efficiency of Mink Oocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3265-3277.
[2]	GUO Yanyan, ZHANG Yuxin, LU Rui, LI Yupeng, CHEN Longbin, ZHANG Jinlong, YAO Dawei, RUAN Weibin, ZHANG Xiaosheng, GUO Xiaofei. Research Progress on the Proliferation and Differentiation of Granulosa Cells at Various Follicular Development Stages in Mammal [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1484-1493.
[3]	YAN Rui, JIA Chaoyang, MA Jing, YANG Juan, LIU Xinfeng, CHEN Qiang. The Research Status and Application Prospect of 3D Culture in Livestock Oocytes and Embryos Culture [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1494-1507.
[4]	SUN Yawen, CHEN Siying, LI Kang, LENG Xuan, WANG Dong, PANG Yunwei. Strategies for Alleviating Cryoinjury of Porcine Vitrified-Oocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 36-44.
[5]	YANG Baigao, XU Jiehuan, ZHANG Liang, LONG Xi, DAI Jianjun, ZHAO Xueming, PAN Hongmei. Exploring the Effect of Vitrification on Genome Methylation Level of Porcine Parthenogenetic Activation Blastocysts by scWGBS [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 222-231.
[6]	Baigao YANG, Xi LONG, Liang ZHANG, Jiehuan XU, Jianjun DAI, Xueming ZHAO, Hongmei PAN. Exploring the Effect of Vitrification on Gene Expression in Porcine Parthenogenetic Blastocysts by Smart-seq2 [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 3936-3946.
[7]	LI Wanjun, XU Jiehuan, HE Mengxian, KONG Yuting, ZHANG Defu, DAI Jianjun. Cytochalasin B Alleviates the Migration Disorder of Cortical Particle Caused by Vitrification in Porcine Oocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 1999-2010.
[8]	WU Zihao, CAI Yilong, TUO Haixin, CHEN Wei. Pathogenicity Analysis of a PVL⁺ ST22 Staphylococcus aureus Isolated from Equine Raw Milk [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 718-726.
[9]	Jianhua DONG, Xiaoyi FENG, Baigao YANG, Chongyang LI, Hongmei PAN, Lihua LÜ, Xueming ZHAO. Advances in Cryopreservation of Porcine Embryo [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 4796-4807.
[10]	SHEN Yingchao, DAVSHILT Toli, REN Hong, WANG Xisheng, TIAN Shuyue, DU Ming, DUGARJAVIIN Manglai, BOU Gerelchimeg. Differential Expression of Oocyte Development-related Hormone and Growth Factor Receptors in Equine Expanded and Compact Cumulus-oocyte Complexes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3735-3744.
[11]	XU Xi, YANG Baigao, ZHANG Hang, FENG Xiaoyi, HAO Haisheng, DU Weihua, ZHU Huabin, ZHANG Peipei, ZHAO Xueming. Effects of NMN on Lipid Droplet Content and Cryopreservation Effect of Bovine Oocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3348-3357.
[12]	YANG Sha, YANG Yuze, XU Xi, HAO Haisheng, DU Weihua, PANG Yunwei, ZHAO Shanjiang, ZOU Huiying, ZHU Huabin, ZHAO Xueming. The Regulation of Methylation Level of IGF2R Gene in IVF Blastocysts Derived from Vitrified Bovine Oocytes by dCas9-SunTag-DNMT3A Technology [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 2015-2023.
[13]	SUN Huan, SUN Kejia, JIANG Xiaolong, LIU Aiju, MA Xiaofei, HAN Hongye, YAO Dawei, MA Yi, TIAN Shujun. Effect of Deoxynivalenol(DON) on Maturation and Development of Bovine Oocytes in vitro [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(9): 2475-2483.
[14]	SHEN Yingchao, REN Hong, NARIGA, WANG Xisheng, MANG Lai, BOU Gerelchimeg. Research Progress of In Vitro Maturation in Horse Oocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 914-922.
[15]	ZHANG Peipei, WANG Jingjing, HAO Haisheng, DU Weihua, PANG Yunwei, QUAN Guobo, ZHAO Shanjiang, ZOU Huiying, HAO Tong, ZHU Huabin, ZHAO Xueming. Study on Whole Genome Methylation Pattern in Vitrified Bovine GV Oocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 1030-1039.