低温保存技术在生物制品中的研究进展

doi:10.19586/j.2095-2341.2023.0045

生物技术进展 ›› 2023, Vol. 13 ›› Issue (4): 547-555.DOI: 10.19586/j.2095-2341.2023.0045

• 进展评述 • 上一篇

低温保存技术在生物制品中的研究进展

刘容麟¹(), 王宁², 李岩异¹(), 张卫婷¹

^1.华北制药金坦生物技术股份有限公司，石家庄 050035
^2.石家庄农业技术推广中心，石家庄 050000

收稿日期:2023-04-04 接受日期:2023-04-28 出版日期:2023-07-25 发布日期:2023-08-03
通讯作者: 李岩异
作者简介:刘容麟 E-mail: liuronglin6@163.com；

Reasearch Progress of Cryopreservation Technology in Biological Products

Ronglin LIU¹(), Ning WANG², Yanyi LI¹(), Weiting ZHANG¹

^1.North China Pharmaceutical Jintan Biotechnology Co. ?, Ltd, Shijiazhuang 050035，China
^2.Shijiazhuang Agricultural Technology Extension Center，Shijiazhuang 050000，China

Received:2023-04-04 Accepted:2023-04-28 Online:2023-07-25 Published:2023-08-03
Contact: Yanyi LI

摘要/Abstract

摘要：

生物制品在储存中易发生多种物理和化学降解，因此生物制品长期保存技术对于减缓生物制品降解、保持生物活性、延长储存期限十分必要。目前，低温保存技术是生物制品长期保存最常用且有效的方法，其能极大地减少生物制品储存时，由于液态水带来的多种物理降解和化学降解导致的活性降低现象。然而低温保存技术在生物制品的保存过程中仍会发生损伤，这些损伤包括冰晶损害、冰水界面吸附变性、渗透伤害等，使用合适的冷冻工艺和冷冻保护剂（cryoprotectant agents, CPAs）对减少低温保存过程带来的负面影响有重要作用。综述了低温保存技术在生物制品保存过程中的研究进展，包括低温保存的影响因素（冷冻保护剂、冷冻过程和其他影响因素）以及最新的低温保存技术，以期为生物制品在长期保存过程中克服损伤和维持活性提供参考。

关键词: 冷冻, 低温保存, 冷冻保护剂, 冻融

Abstract:

Biological products are prone to various physical and chemical degradation during storage， so long-term storage technology for biological products is necessary to slow down degradation， maintain biological activity and extend storage life. At present， low-temperature preservation technology is the most commonly used and effective method for long-term preservation of biological products， which can greatly reduce the activity decrease resulted from various physical and chemical degradation caused by liquid water during the storage of biological products. However， there are still some damages in the preservation process of biological products using low-temperature preservation technology， such as ice crystal damage， adsorption denaturation of ice water interface， and osmotic damage. The use of appropriate freezing processes and cryoprotectant agents （CPAs） plays an important role in reducing the negative impact of low-temperature preservation process. This article reviewed the research progress of low-temperature preservation technology in the preservation of biological products， including the influencing factors of low-temperature preservation （cryoprotectants， freezing processes and other influencing factors）， as well as the latest low-temperature preservation technologies， in order to provide reference for overcoming damage and maintaining activity of biological products during long-term preservation.

Key words: freeze, cryopreservation, cryoprotectant, freeze thawing

中图分类号:

TQ464

刘容麟, 王宁, 李岩异, 张卫婷. 低温保存技术在生物制品中的研究进展[J]. 生物技术进展, 2023, 13(4): 547-555.

Ronglin LIU, Ning WANG, Yanyi LI, Weiting ZHANG. Reasearch Progress of Cryopreservation Technology in Biological Products[J]. Current Biotechnology, 2023, 13(4): 547-555.

图/表 2

参考文献 46

1	MAZUR P. Cryobiology: the freezing of biological systems[J]. Science, 1970, 168(3934): 939-949.
2	FARRANT J. Mechanism of cell damage during freezing and thawing and its prevention[J]. Nature,1965,205(4978):1284-1287.
3	OLDENHOF H, WOLKERS W F, SIEME H. Cryopreservation of Semen from domestic livestock: bovine, equine, and porcine sperm[J]. Methods Mol. Biol., 2021, 2180: 365-377.
4	MARSOLA S J, JORGE L F, MENIQUETI A B, et al.. Endophytic fungi of Brunfelsia uniflora: isolation, cryopreservation, and determination of enzymatic and antioxidant activity[J/OL]. World J. Microbiol. Biotechnol.,2022,38:94[2022-04-21]. .
5	POLGE C, SMITH A U, PARKES A S. Revival of spermatozoa after vitrification and dehydration at low temperatures[J/OL]. Nature, 1949, 164(4172): 666[1949-10-15]. .
6	LOVELOCK J E, BISHOP M W. Prevention of freezing damage to living cells by dimethyl sulphoxide[J]. Nature, 1959, 183(4672): 1394-1395.
7	MOKOENA K, SYM S, MYCOCK D. Cryopreservation of Pyramimonas mucifera [J]. Cryoletters, 2022, 43(1): 18-24.
8	PRIETO-GUEVARA M, ALARCÓN-FURNIELES J, JIMÉNEZ-VELÁSQUEZ C, et al.. Cryopreservation of the microalgae Scenedesmus sp.[J/OL]. Cells,2023,12: 562[2023-02-09]. .
9	NIKITIN G, MAKAVCHIK S, SUKHININ A, et al.. Aspects of Mannheimia haemolytica cryopreservation[J/OL]. FASEB J.,2022,36: R3117[2022-05-01]. .
10	KARDORFF M, MAHLER H C, HUWYLER J, et al.. Cryoprotection in human mesenchymal stromal/stem cells: synergistic impact of urea and glucose[J]. J. Pharm. Sci.,2023, 6: 1681-1686.
11	CHAN A, MATURANA C J, ENGEL E A. Optimized formulation buffer preserves adeno-associated virus-9 infectivity after 4 ℃ storage and freeze/thawing cycling[J/OL]. J. Virol. Methods, 2022, 309: 114598[2022-08-05]. .
12	MURRAY K A, GIBSON M I. Chemical approaches to cryopreservation[J]. Nat. Rev. Chem., 2022, 6(8): 579-593.
13	MARTON H L, STYLES K M, KILBRIDE P, et al.. Polymer-mediated cryopreservation of bacteriophages[J]. Biomacromolecules, 2021, 22(12): 5281-5289.
14	MITCHELL D E, LOVETT J R, ARMES S P, et al.. Combining biomimetic block copolymer worms with an ice-inhibiting polymer for the solvent-free cryopreservation of red blood cells[J]. Angew. Chem. Int. Ed. Engl., 2016, 55(8): 2801-2804.
15	FUJITA Y, NISHIMURA M, WADA T, et al.. Dimethyl sulfoxide-free cryopreservation solution containing trehalose, dextran 40, and propylene glycol for therapy with human adipose tissue-derived mesenchymal stromal cells[J]. Cytotechnology, 2022, 74(5): 515-529.
16	HAUPTMANN A, PODGORSEK K, KUZMAN D, et al.. Impact of buffer, protein concentration and sucrose addition on the aggregation and particle formation during freezing and thawing[J/OL]. Pharm. Res., 2018, 35: 101[2018-03-21]. .
17	SRETER J A, FOXALL T L, VARGA K. Intracellular and extracellular antifreeze protein significantly improves mammalian cell cryopreservation[J/OL]. Biomolecules,2022,12: 669[2022-05-29]. .
18	SUN Y, MALTSEVA D, LIU J, et al.. Ice recrystallization inhibition is insufficient to explain cryopreservation abilities of antifreeze proteins[J]. Biomacromolecules, 2022, 23(3): 1214-1220.
19	THORAT A A, MUNJAL B, GEDERS T W, et al.. Freezing-induced protein aggregation-role of pH shift and potential mitigation strategies[J]. J. Control. Release, 2020, 323: 591-599.
20	REDA ELKHAWAGAH A, MARTINO N A, MOUSTAFA KANDIEL M M, et al.. Effects of cysteamine supplementation on cryopreserved buffalo bull Semen quality parameters[J]. Theriogenology, 2022, 192: 141-149.
21	FAYAZI S, DAMVAR N, MOLAEIAN S, et al.. Thermally conductive graphene-based nanofluids, a novel class of cryosolutions for mouse blastocysts vitrification[J/OL]. Reprod. Biol., 2022, 22: 100635[2022-06-01]. .
22	FEYZMANESH S, HALVAEI I, BAHEIRAEI N. Alginate effects on human sperm parameters during freezing and thawing: a prospective study[J]. Cell J., 2022, 24(7): 417-423.
23	DIAZ-DUSSAN D, PENG Y Y, SENGUPTA J, et al.. Trehalose-based polyethers for cryopreservation and three-dimensional cell scaffolds[J]. Biomacromolecules, 2020, 21(3): 1264-1273.
24	CHANG T, ZHAO G. Ice inhibition for cryopreservation: materials, strategies, and challenges[J/OL]. Adv. Sci.,2021,8:202002425[2021-03-17]. .
25	PARK S, PIAO Z, PARK J K, et al.. Ice recrystallization inhibition using L-alanine/L-lysine copolymers[J]. ACS Appl. Polym. Mater., 2022, 4(4): 2896-2907.
26	BOAFO G F, MAGAR K T, EKPO M D, et al.. The role of cryoprotective agents in liposome stabilization and preservation[J/OL]. Int. J. Mol. Sci., 2022, 23: 12487[2022-10-28]. .
27	GUO N, SONG Y, YAN J, et al.. The effect of cryopreservation on the survival of Nocardia farcinica and Yersinia pestis vaccine strains[J/OL]. Biopreserv. Biobank., 2022: 0049[2022-09-20]. .
28	XIN Y, KIELAR C, ZHU S, et al.. Cryopreservation of DNA origami nanostructures[J/OL]. Small, 2020, 16: e1905959[2020-03-05]. .
29	NAULLAGE P M, MOLINERO V. Slow propagation of ice binding limits the ice-recrystallization inhibition efficiency of PVA and other flexible polymers[J]. J. Am. Chem. Soc., 2020, 142(9): 4356-4366.
30	QIN Q, ZHAO L, LIU Z, et al.. Bioinspired L-proline oligomers for the cryopreservation of oocytes via controlling ice growth[J]. ACS Appl. Mater. Interfaces, 2020, 12(16): 18352-18362.
31	SINGH P, BEDI M K, SINGHAL S, et al.. Effect of graphene oxide as cryoprotectant on post-thaw sperm functional and kinetic parameters of cross bred (HF X Sahiwal) and Murrah buffalo (Bubalus bubalis) bulls[J]. Cryobiology, 2022, 106: 102-112.
32	CAO R, SOGABE T, MIKAJIRI S, et al.. Effects of sucrose, carnosine, and their mixture on the glass transition behavior and storage stability of freeze-dried lactic acid bacteria at various water activities[J]. Cryobiology, 2022, 106: 131-138.
33	XU Y, LI-YING C L, CHEN S, et al.. Optimization of UC-MSCs cold-chain storage by minimizing temperature fluctuations using an automatic cryopreservation system[J]. Cryobiology, 2021, 99: 131-139.
34	MCGUINNESS S, CHARLEWOOD R, GILDER E, et al.. A pilot randomized clinical trial of cryopreserved versus liquid-stored platelet transfusion for bleeding in cardiac surgery: the cryopreserved versus liquid platelet-New Zealand pilot trial[J]. Vox Sang., 2022, 117(3): 337-345.
35	ZHAN L, LI M G, HAYS T, et al.. Cryopreservation method for Drosophila melanogaster embryos[J/OL]. Nat. Commun., 2021, 12(1): 2412[2021-04-23]. .
36	ZHANG P Q, TAN P C, GAO Y M, et al.. The effect of glycerol as a cryoprotective agent in the cryopreservation of adipose tissue[J/OL]. Stem Cell Res. Ther.,2022,13:152[2022-04-08]..
37	CAO E, CHEN Y, CUI Z, et al.. Effect of freezing and thawing rates on denaturation of proteins in aqueous solutions[J]. Biotechnol. Bioeng., 2003, 82(6): 684-690.
38	MINATOVICZ B, SUN L, FORAN C, et al.. Freeze-concentration of solutes during bulk freezing and its impact on protein stability[J/OL]. J. Drug Deliv. Sci. Technol., 2020, 58: 101703 [2021-04-01]. .
39	ZHANG Y, ERTBJERG P. On the origin of thaw loss: Relationship between freezing rate and protein denaturation[J/OL]. Food Chem.,2019,299:125104[2019-07-07]. .
40	SHIKAMA K. Effect of freezing and thawing on the stability of double helix of DNA[J]. Nature, 1965, 207(996): 529-530.
41	FUCHIZAKI A, YASUI K, TANAKA M, et al.. Comparison of the programmed freezer method and deep freezer method in the manufacturing of frozen red blood cell products[J]. Vox Sang., 2022, 117(6): 812-821.
42	JAMES E R, WEN Y, OVERBY J, et al.. Cryopreservation of Anopheles stephensi embryos[J/OL]. Sci. Rep., 2022, 12(1): 43[2022-01-07]. .
43	LI X, SIVIROJ P, RUANGSURIYA J, et al.. Effects of the thawing rate and heating temperature on immunoglobulin A and lysozyme activity in human milk[J/OL]. Int. Breastfeed. J., 2022, 17(1): 52[2022-07-07]. .
44	COTTLE C, PORTER A P, LIPAT A, et al.. Impact of cryopreservation and freeze-thawing on therapeutic properties of mesenchymal stromal/stem cells and other common cellular therapeutics[J]. Curr. Stem Cell Rep., 2022, 8(2): 72-92.
45	AGARWAL A, ROSAS I M, ANAGNOSTOPOULOU C, et al.. Oxidative stress and assisted reproduction: a comprehensive review of its pathophysiological role and strategies for optimizing embryo culture environment[J/OL]. Antioxidants Basel.,2022,11: 22[2022-02-28]. .
46	MA Y, GAO L, TIAN Y, et al.. Advanced biomaterials in cell preservation: hypothermic preservation and cryopreservation[J]. Acta Biomater., 2021, 131: 97-116.

保护剂渗透类型	种类	名称	保护作用	参考文献
渗透型	有机小分子	甘油	渗透保护、抑制冰晶	[5]
渗透型	有机小分子	二甲基亚砜	渗透保护、抑制冰晶	[7]
渗透型	有机小分子	乙二醇	渗透保护、抑制冰晶	[9]
渗透型	有机小分子	丙二醇	渗透保护、抑制冰晶	[15]
甲醇非渗透型	糖	蔗糖	抑制胞外大冰晶、提高玻璃转化温度	[16]
非渗透型	糖	海藻糖	抑制胞外大冰晶、提高玻璃转化温度	[15]
非渗透型	糖	甘露醇	抑制胞外大冰晶、提高玻璃转化温度	[16]
非渗透型	糖	葡萄糖	抑制胞外大冰晶	[10]
非渗透型	聚合物	羟乙基淀粉	抑制胞外大冰晶	[12]
非渗透型	蛋白质	抗冻蛋白	抑制胞外大冰晶、抑制冰重结晶	[17]
非渗透型	糖蛋白	抗冻糖蛋白	抑制胞外大冰晶、抑制冰重结晶	[18]
非渗透型	蛋白质	白蛋白	抑制胞外大冰晶	[19]
非渗透型	氨基酸	半胱氨酸	抗氧化	[20]
非渗透型	纳米材料	氧化石墨烯	封装保护，快速导热	[21]
非渗透型	聚合物	海藻酸盐	包封保护	[22]
非渗透型	聚合物	聚乙二醇	包封保护	[22]
非渗透型	聚合物	海藻糖基聚合物	包封保护	[23]
非渗透型	聚合物	水凝胶	包封保护生物材料、抑制胞外大冰晶	[24]
非渗透型	聚合物	聚乙烯醇	抑制冰重结晶、影响冰形状	[14]
非渗透型	聚合物	聚脯氨酸	包封保护生物材料、抑制冰晶、抑制重结晶	[14]
非渗透型	聚合物	聚(L-丙氨酸-co-L-赖氨酸)	抑制冰重结晶、影响冰形状	[25]

保护剂渗透类型	种类	名称	保护作用	参考文献
渗透型	有机小分子	甘油	渗透保护、抑制冰晶	[5]
渗透型	有机小分子	二甲基亚砜	渗透保护、抑制冰晶	[7]
渗透型	有机小分子	乙二醇	渗透保护、抑制冰晶	[9]
渗透型	有机小分子	丙二醇	渗透保护、抑制冰晶	[15]
甲醇非渗透型	糖	蔗糖	抑制胞外大冰晶、提高玻璃转化温度	[16]
非渗透型	糖	海藻糖	抑制胞外大冰晶、提高玻璃转化温度	[15]
非渗透型	糖	甘露醇	抑制胞外大冰晶、提高玻璃转化温度	[16]
非渗透型	糖	葡萄糖	抑制胞外大冰晶	[10]
非渗透型	聚合物	羟乙基淀粉	抑制胞外大冰晶	[12]
非渗透型	蛋白质	抗冻蛋白	抑制胞外大冰晶、抑制冰重结晶	[17]
非渗透型	糖蛋白	抗冻糖蛋白	抑制胞外大冰晶、抑制冰重结晶	[18]
非渗透型	蛋白质	白蛋白	抑制胞外大冰晶	[19]
非渗透型	氨基酸	半胱氨酸	抗氧化	[20]
非渗透型	纳米材料	氧化石墨烯	封装保护，快速导热	[21]
非渗透型	聚合物	海藻酸盐	包封保护	[22]
非渗透型	聚合物	聚乙二醇	包封保护	[22]
非渗透型	聚合物	海藻糖基聚合物	包封保护	[23]
非渗透型	聚合物	水凝胶	包封保护生物材料、抑制胞外大冰晶	[24]
非渗透型	聚合物	聚乙烯醇	抑制冰重结晶、影响冰形状	[14]
非渗透型	聚合物	聚脯氨酸	包封保护生物材料、抑制冰晶、抑制重结晶	[14]
非渗透型	聚合物	聚(L-丙氨酸-co-L-赖氨酸)	抑制冰重结晶、影响冰形状	[25]

低温保存技术在生物制品中的研究进展

Reasearch Progress of Cryopreservation Technology in Biological Products

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 2

参考文献 46

相关文章 1

编辑推荐

Metrics

本文评价