猪卵母细胞玻璃化冷冻损伤的缓解策略

doi:10.11843/j.issn.0366-6964.2025.01.004

摘要/Abstract

摘要：

卵母细胞的冷冻保存是人类和动物辅助生殖中长期储存遗传资源的重要策略之一。然而，由于卵母细胞的表面积/体积比小、细胞渗透性差，在冷冻过程中极易受到冰晶形成、渗透性休克、氧化应激和冷冻保护剂毒性等损伤，严重降低其解冻后的发育能力。与其他动物相比，猪卵母细胞中丰富的脂滴含量降低了其低温耐受性，冷冻保存难度更大。尽管可以从玻璃化未成熟卵母细胞中获得活仔猪，但与新鲜卵母细胞相比，玻璃化冷冻卵母细胞的发育能力还有待进一步提升。本文主要从玻璃化冷冻猪卵母细胞的损伤类型、受损机制、缓解冷冻损伤的策略等方面进行综述，为进一步建立和完善猪卵母细胞玻璃化冷冻保存方案提供参考。

关键词: 猪, 卵母细胞, 玻璃化冷冻, 冷冻损伤

Abstract:

Cryopreservation of oocytes is one of the important strategies for long-term storage of genetic resources in both humans and animal-assisted reproduction. However, due to the low surface-volume ratio and poor cell permeability, oocytes are more susceptible to cryoinjuries during vitrification, including ice crystals formation, osmotic shock, oxidative stress, and cryoprotectants toxicity, which seriously decreases their developmental ability after thawing. In contrast to other animals, porcine oocyte cryopreservation is more challenging owing to its abundant lipid droplets and poor hypothermia tolerance. Although live piglets have been produced from vitrified immature oocytes, the developmental competence of vitrified-oocytes remains to be further promoted compared with fresh oocytes. This paper mainly reviews the injury types, the mechanisms of damage, as well as the alleviating strategies of porcine vitrified-oocytes. It will provide further reference for establishing and improving the vitrification protocol of porcine oocytes.

Key words: porcine, oocytes, vitrification, cryoinjury

中图分类号:

S814.6

孙雅雯, 陈思颍, 李伉, 冷璇, 王栋, 庞云渭. 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.

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.

图/表 2

参考文献 74

1	SOMFAI T . Vitrification of immature oocytes in pigs[J]. Anim Sci J, 2024, 95 (1): e13943. doi: 10.1111/asj.13943
2	MCEVOY T G , COULL G D , BROADBENT P J , et al. Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida[J]. J Reprod Fertil, 2000, 118 (1): 163- 170. doi: 10.1530/reprod/118.1.163
3	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
4	SOMFAI T , YOSHIOKA K , TANIHARA F , et al. Generation of live piglets from cryopreserved oocytes for the first time using a defined system for in vitro embryo production[J]. PLoS One, 2014, 9 (5): e97731. doi: 10.1371/journal.pone.0097731
5	AMINI M , BENSON J D . Technologies for vitrification based cryopreservation[J]. Bioengineering (Basel), 2023, 10 (5): 508. doi: 10.3390/bioengineering10050508
6	ZHU Y X , LIU H Y , ZHENG L , et al. Vitrification of mammalian oocytes: recent studies on mitochondrial dysfunction[J]. Biopreserv Biobank, 2024, 22 (5): 428- 440. doi: 10.1089/bio.2023.0062
7	LIU R H , SUN Q Y , LI Y H , et al. Effects of cooling on meiotic spindle structure and chromosome alignment within in vitro matured porcine oocytes[J]. Mol Reprod Dev, 2003, 65 (2): 212- 218. doi: 10.1002/mrd.10282
8	CASILLAS F , DUCOLOMB Y , LÓPEZ A , et al. Effect of porcine immature oocyte vitrification on oocyte-cumulus cell gap junctional intercellular communication[J]. Porcine Health Manag, 2020, 6 (1): 37. doi: 10.1186/s40813-020-00175-x
9	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
10	WU C H , RUI R , DAI J J , et al. Effects of cryopreservation on the developmental competence, ultrastructure and cytoskeletal structure of porcine oocytes[J]. Mol Reprod Dev, 2006, 73 (11): 1454- 1462. doi: 10.1002/mrd.20579
11	周悦, 卢俊求, 吴亚辉, 等. 玻璃化冷冻对猪GV期卵母细胞成熟及发育能力的影响[J]. 中国畜牧杂志, 2015, 51 (5): 29- 33. doi: 10.3969/j.issn.0258-7033.2015.05.006
	ZHOU Y , LU J Q , WU Y H , et al. Effects of vitrification on in vitro development ability of porcine oocytes at GV stage[J]. Chinese Journal of Animal Science, 2015, 51 (5): 29- 33. doi: 10.3969/j.issn.0258-7033.2015.05.006
12	张超凡, 薛玉环, 左芙蓉, 等. 玻璃化冷冻对小鼠卵母细胞超微结构的影响[J]. 畜牧与兽医, 2017, 49 (6): 72- 76.
	ZHANG C F , XUE Y H , ZUO F R , et al. Effects of vitrification on ultrastructure of mouse oocytes[J]. Animal Husbandry & Veterinary Medicine, 2017, 49 (6): 72- 76.
13	OKOTRUB K A , OMELCHENKO A N , CHUYKO E A , et al. Irreversible lipid phase transition detected in a porcine oocyte at chilling[J]. Cryobiology, 2024, 114, 104850. doi: 10.1016/j.cryobiol.2024.104850
14	MA Y M , GAO L , TIAN Y Q , et al. Advanced biomaterials in cell preservation: hypothermic preservation and cryopreservation[J]. Acta Biomater, 2021, 131, 97- 116. doi: 10.1016/j.actbio.2021.07.001
15	SOMFAI T , HARAGUCHI S , DANG-NGUYEN T Q , et al. Vitrification of porcine immature oocytes and zygotes results in different levels of DNA damage which reflects developmental competence to the blastocyst stage[J]. PLoS One, 2023, 18 (3): e0282959. doi: 10.1371/journal.pone.0282959
16	MATEO-OTERO Y , YESTE M , DAMATO A , et al. Cryopreservation and oxidative stress in porcine oocytes[J]. Res Vet Sci, 2021, 135, 20- 26. doi: 10.1016/j.rvsc.2020.12.024
17	DAI J J , WU C F , MUNERI C W , et al. Changes in mitochondrial function in porcine vitrified MⅡ-stage oocytes and their impacts on apoptosis and developmental ability[J]. Cryobiology, 2015, 71 (2): 291- 298. doi: 10.1016/j.cryobiol.2015.08.002
18	DAI J J , NIU Y F , WU C F , et al. Both death receptor and mitochondria mediated apoptotic pathways participated the occurrence of apoptosis in porcine vitrified MⅡ stage oocytes[J]. Cryo Letters, 2016, 37 (2): 129- 136.
19	DAI J J , YANG J H , ZHANG S S , et al. Partial recovery of mitochondrial function of vitrified porcine MⅡ stage oocytes during post-thaw incubation[J]. Cryo Letters, 2018, 39 (1): 39- 44.
20	CAO B J , QIN J P , PAN B , et al. Oxidative stress and oocyte cryopreservation: recent advances in mitigation strategies involving antioxidants[J]. Cells, 2022, 11 (22): 3573. doi: 10.3390/cells11223573
21	JIA B Y , XIANG D C , YANG H , et al. Transcriptome analysis of porcine embryos derived from oocytes vitrified at the germinal vesicle stage[J]. Theriogenology, 2024, 218, 99- 110. doi: 10.1016/j.theriogenology.2024.01.032
22	JIA B Y , XIANG D C , ZHANG B , et al. Quality of vitrified porcine immature oocytes is improved by coculture with fresh oocytes during in vitro maturation[J]. Mol Reprod Dev, 2019, 86 (11): 1615- 1627. doi: 10.1002/mrd.23249
23	ITO J , SHIRASUNA K , KUWAYAMA T , et al. Resveratrol treatment increases mitochondrial biogenesis and improves viability of porcine germinal-vesicle stage vitrified-warmed oocytes[J]. Cryobiology, 2020, 93, 37- 43. doi: 10.1016/j.cryobiol.2020.02.014
24	SCIORIO R , CAMPOS G , TRAMONTANO L , et al. Exploring the effect of cryopreservation in assisted reproductive technology and potential epigenetic risk[J]. Zygote, 2023, 31 (5): 420- 432. doi: 10.1017/S0967199423000345
25	SCIORIO R , MANNA C , FAUQUE P , et al. Can cryopreservation in assisted reproductive technology (ART) induce epigenetic changes to gametes and embryos?[J]. J Clin Med, 2023, 12 (13): 4444. doi: 10.3390/jcm12134444
26	CHENG K R , FU X W , ZHANG R N , et al. Effect of oocyte vitrification on deoxyribonucleic acid methylation of H19, Peg3, and Snrpn differentially methylated regions in mouse blastocysts[J]. Fertil Steril, 2014, 102 (4): 1183- 1190. doi: 10.1016/j.fertnstert.2014.06.037
27	MOULAVI F , SAADELDIN I M , SWELUM A A , et al. Oocyte vitrification induces loss of DNA methylation and histone acetylation in the resulting embryos derived using ICSI in dromedary camel[J]. Zygote, 2021, 29 (5): 383- 392. doi: 10.1017/S0967199421000150
28	MA Y Z , LONG C S , 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
29	SPINACI M , VALLORANI C , BUCCI D , et al. Vitrification of pig oocytes induces changes in histone H4 acetylation and histone H3 lysine 9 methylation (H3K9)[J]. Vet Res Commun, 2012, 36 (3): 165- 171. doi: 10.1007/s11259-012-9527-9
30	CHEN H H , ZHANG L , DENG T F , 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
31	SHADMANESH A , NAZARI H . Alterations in the expression pattern of some epigenetic-related genes and microRNAs subsequent to oocyte cryo-preservation[J]. Zygote, 2023, 31 (5): 411- 419. doi: 10.1017/S0967199423000321
32	JIA B Y , XIANG D C , QUAN G B , et al. Transcriptome analysis of porcine immature oocytes and surrounding cumulus cells after vitrification and in vitro maturation[J]. Theriogenology, 2019, 134, 90- 97. doi: 10.1016/j.theriogenology.2019.05.019
33	BARBERET J , BARRY F , CHOUX C , et al. What impact does oocyte vitrification have on epigenetics and gene expression?[J]. Clin Epigenetics, 2020, 12 (1): 121. doi: 10.1186/s13148-020-00911-8
34	MURRAY K A , GIBSON M I . Chemical approaches to cryopreservation[J]. Nat Rev Chem, 2022, 6 (8): 579- 593. doi: 10.1038/s41570-022-00407-4
35	CHANG C C , SHAPIRO D B , NAGY Z P . The effects of vitrification on oocyte quality[J]. Biol Reprod, 2022, 106 (2): 316- 327. doi: 10.1093/biolre/ioab239
36	LIU Z , ZHENG X , WANG J J . Bioinspired ice-binding materials for tissue and organ cryopreservation[J]. J Am Chem Soc, 2022, 144 (13): 5685- 5701. doi: 10.1021/jacs.2c00203
37	WIESAK T , WASIELAK M , ZŁOTKOWSKA A , et al. Effect of vitrification on the zona pellucida hardening and follistatin and cathepsin B genes expression and developmental competence of in vitro matured bovine oocytes[J]. Cryobiology, 2017, 76, 18- 23. doi: 10.1016/j.cryobiol.2017.05.001
38	RUSCIANO G , DE CANDITⅡS C , ZITO G , et al. Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes[J]. PLoS One, 2017, 12 (5): e0177677. doi: 10.1371/journal.pone.0177677
39	CHEN J M , LIU X J , HU Y Y , et al. Cryopreservation of tissues and organs: present, bottlenecks, and future[J]. Front Vet Sci, 2023, 10, 1201794. doi: 10.3389/fvets.2023.1201794
40	WU G Q , JIA B Y , QUAN G B , et al. Vitrification of porcine immature oocytes: association of equilibration manners with warming procedures, and permeating cryoprotectants effects under two temperatures[J]. Cryobiology, 2017, 75, 21- 27. doi: 10.1016/j.cryobiol.2017.03.001
41	ARAV A , ZERON Y , LESLIE S B , et al. Phase transition temperature and chilling sensitivity of bovine oocytes[J]. Cryobiology, 1996, 33 (6): 589- 599. doi: 10.1006/cryo.1996.0062
42	DOU M J , LU C N , RAO W . Bioinspired materials and technology for advanced cryopreservation[J]. Trends Biotechnol, 2022, 40 (1): 93- 106. doi: 10.1016/j.tibtech.2021.06.004
43	TANG Y , ZHANG Y , LIU L X , et al. Glycine and melatonin improve preimplantation development of porcine oocytes vitrified at the germinal vesicle stage[J]. Front Cell Dev Biol, 2022, 10, 856486. doi: 10.3389/fcell.2022.856486
44	PELUSO G , BARBARISI A , SAVICA V , et al. Carnitine: an osmolyte that plays a metabolic role[J]. J Cell Biochem, 2000, 80 (1): 1- 10.
45	GORE M , NARVEKAR A , BHAGWAT A , et al. Macromolecular cryoprotectants for the preservation of mammalian cell culture: lessons from crowding, overview and perspectives[J]. J Mater Chem B, 2022, 10 (2): 143- 169. doi: 10.1039/D1TB01449H
46	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 X , 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
47	MORATÓ R , CHAUVIGNÉ F , NOVO S , et al. Enhanced water and cryoprotectant permeability of porcine oocytes after artificial expression of human and zebrafish aquaporin-3 channels[J]. Mol Reprod Dev, 2014, 81 (5): 450- 461. doi: 10.1002/mrd.22310
48	DRORI R , STEVENS C A . Divergent mechanisms of ice growth inhibition by antifreeze proteins[J]. Methods Mol Biol, 2024, 2730, 169- 181.
49	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.
50	SANTOS E C D S , SOMFAI T , APPELTANT R , et al. Effects of polyethylene glycol and a synthetic ice blocker during vitrification of immature porcine oocytes on survival and subsequent embryo development[J]. Anim Sci J, 2017, 88 (8): 1042- 1048. doi: 10.1111/asj.12730
51	KAMOSHITA M , KATO T , FUJIWARA K , et al. Successful vitrification of pronuclear-stage pig embryos with a novel cryoprotective agent, carboxylated ε-poly-L-lysine[J]. PLoS One, 2017, 12 (4): e0176711. doi: 10.1371/journal.pone.0176711
52	KAWASAKI Y , KOHAYA N , SHIBAO Y , et al. Carboxylated ε-poly-L-lysine, a cryoprotective agent, is an effective partner of ethylene glycol for the vitrification of embryos at various preimplantation stages[J]. Cryobiology, 2020, 97, 245- 249. doi: 10.1016/j.cryobiol.2020.10.004
53	李维杰, 周新丽, 刘宝林, 等. 纳米颗粒对猪GV期卵母细胞低温保存效果的影响[J]. 中国生物医学工程学报, 2013, 32 (5): 601- 605.
	LI W J , ZHOU X L , LIU B L , et al. The Effect of nanoparticle on vitrification of porcine GV-stage oocytes[J]. Chinese Journal of Biomedical Engineering, 2013, 32 (5): 601- 605.
54	MASSIE I , SELDEN C , HODGSON H , et al. Cryopreservation of encapsulated liver spheroids for a bioartificial liver: reducing latent cryoinjury using an ice nucleating agent[J]. Tissue Eng Part C Methods, 2011, 17 (7): 765- 774. doi: 10.1089/ten.tec.2010.0394
55	CROCKETT E L . The cold but not hard fats in ectotherms: consequences of lipid restructuring on susceptibility of biological membranes to per-oxidation, a review[J]. J Comp Physiol B, 2008, 178 (7): 795- 809. doi: 10.1007/s00360-008-0275-7
56	BARRERO-SICILIA C , SILVESTRE S , HASLAM R P , et al. Lipid remodelling: unravelling the response to cold stress in Arabidopsis and its extremophile relative Eutrema salsugineum[J]. Plant Sci, 2017, 263, 194- 200. doi: 10.1016/j.plantsci.2017.07.017
57	IBAYASHI M , TSUKAMOTO S . Lipid remodelling in mammalian development[J]. Nat Cell Biol, 2024, 26 (2): 179- 180. doi: 10.1038/s41556-023-01327-1
58	JUNG G T , LEE J H , PARK D , et al. Lipidomic changes in mouse oocytes vitrified in PEG 8000-supplemented vitrification solutions[J]. Cryobiology, 2021, 99, 140- 148. doi: 10.1016/j.cryobiol.2020.11.004
59	AARDEMA H , BERTIJN I , VAN TOL H , et al. Fatty acid supplementation during in vitro embryo production determines cryosurvival characteristics of bovine blastocysts[J]. Front Cell Dev Biol, 2022, 10, 837405. doi: 10.3389/fcell.2022.837405
60	CHEN X , DONG H T , CHENG M M , et al. Addition of cholesterol loaded cyclodextrin prior to GV-phase vitrification improves the quality of mature porcine oocytes in vitro[J]. Cryobiology, 2019, 90, 54- 62. doi: 10.1016/j.cryobiol.2019.08.006
61	FU X W , WU G Q , LI J J , et al. Positive effects of Forskolin (stimulator of lipolysis) treatment on cryosurvival of in vitro matured porcine oocytes[J]. Theriogenology, 2011, 75 (2): 268- 275. doi: 10.1016/j.theriogenology.2010.08.013
62	张紫薇, 于博, 戴佳格, 等. 小檗碱对玻璃化冷冻猪卵母细胞脂滴含量及胚胎发育的影响[J]. 北京农学院学报, 2020, 35 (1): 67- 74.
	ZHANG Z W , YU B , DAI J G , et al. Effects of berberine on lipid droplets content and embryo development in vitrified frozen porcine oocytes[J]. Journal of Beijing University of Agriculture, 2020, 35 (1): 67- 74.
63	XU H X , JIA C , CHENG W X , et al. The effect of L-carnitine additive during in vitro maturation on the vitrification of pig oocytes[J]. Cell Reprogram, 2020, 22 (4): 198- 207. doi: 10.1089/cell.2020.0014
64	蔡绍莉, 徐皆欢, 何孟纤, 等. 毛喉素对猪卵母细胞降脂及冷冻保护效果研究[J]. 畜牧兽医学报, 2023, 54 (1): 178- 188. doi: 10.11843/j.issn.0366-6964.2023.01.017
	CAI S L , XU J H , HE M X , et al. Effects of forskolin on lipid degradation and cryopreservation of porcine oocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54 (1): 178- 188. doi: 10.11843/j.issn.0366-6964.2023.01.017
65	SANTOS E , APPELTANT R , DANG-NGUYEN T Q , et al. The effect of resveratrol on the developmental competence of porcine oocytes vitrified at germinal vesicle stage[J]. Reprod Domest Anim, 2018, 53 (2): 304- 312. doi: 10.1111/rda.13105
66	王文杰, 葛雷, 张树山, 等. 线粒体靶向抗氧化剂MitoQ对猪孤雌激活囊胚冷冻保存的影响[J]. 上海农业学报, 2021, 37 (1): 60- 65.
	WANG W J , GE L , ZHANG S S , et al. Application of mitochondria-targeted antioxidant MitoQ in cryo-preservation of porcine parthenogenetic blastocysts[J]. Acta Agriculturae Shanghai, 2021, 37 (1): 60- 65.
67	XU J H , SUN L W , WU C F , et al. Involvement of PINK1/Parkin-mediated mitophagy in mitochondrial functional disruption under oxidative stress in vitrified porcine oocytes[J]. Theriogenology, 2021, 174, 160- 168.
68	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.
69	NIU Y F , DAI J J , CHEN Y N , et al. Positive effect of apoptotic inhibitor Z-VAD-FMK on vitrified-thawed porcine MⅡ stage oocytes[J]. Cryo Letters, 2016, 37 (3): 188- 195.
70	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
71	GUO Y Y , YANG Y , YI X Y , et al. Microfluidic method reduces osmotic stress injury to oocytes during cryoprotectant addition and removal processes in porcine oocytes[J]. Cryobiology, 2019, 90, 63- 70.
72	周新丽, 郭莹莹, 衣星越, 等. 微流控芯片用于卵母细胞冷冻保存的实验研究[J]. 生物化学与生物物理进展, 2018, 45 (7): 763- 771.
	ZHOU X L , GUO Y Y , YI X Y , et al. Experimental study of microfluidic chip for cryopreservation of oocytes[J]. Progress in Biochemistry and Biophysics, 2018, 45 (7): 763- 771.
73	PRIBENSZKY C , VAJTA G , MOLNAR M , et al. Stress for stress tolerance?A fundamentally new approach in mammalian embryology[J]. Biol Reprod, 2010, 83 (5): 690- 697.
74	DU Y , PRIBENSZKY C S , MOLNÁR M , et al. High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification[J]. Reproduction, 2008, 135 (1): 13- 17.

缓解策略 Alleviating strategy	优点 Advantage	缺点 Disadvantage
增强渗透调节 Enhancement of osmotic regulation	平衡细胞内外渗透压，降低冷冻保护剂毒性	具有浓度依赖性，且物种之间的差异较大
抑制冰晶形成 Inhibition of ice crystals formation	抑制冰晶生长，降低溶液冰点	控冰材料价格相对昂贵，生物相容性较差
促进脂质重塑 Promotion of lipid remodeling	稳定细胞膜组成，维持细胞膜流动性	改变了卵母细胞固有的脂质结构特点，对后续发育的影响需进一步探索
降低氧化损伤 Reduction of oxidative damage	减少ROS生成和凋亡发生率	不同抗氧化剂的使用效果差异较大
工程化策略 Engineering strategy	仿生控冰，对细胞损伤较小	成本较高，可用范围局限

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