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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2022, Vol. 42 Issue (1/2): 119-127    DOI: 10.13523/j.cb.2108059
综述     
定容条件下生物低温保存研究进展*
赵远恒1,2,郭嘉1,陈六彪1,**(),王俊杰1,2
1 中国科学院低温工程学重点实验室 中国科学院理化技术研究所 北京 100190
2 中国科学院大学 北京 100049
A Review on Bio-preservation at Sub-freezing Temperatures under Isochoric Conditions
ZHAO Yuan-heng1,2,GUO Jia1,CHEN Liu-biao1,**(),WANG Jun-jie1,2
1 Key Laboratory of Cryogenics, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190, China
2 University of Chinese Academy of Sciences, Beijing 100049, China
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摘要：

低温能降低生物体内生化反应速率,延长生物活性时间。为了避免传统常压冷冻保存过程中冰晶的产生对生物组织造成的破坏,Dr. Rubinsky提出2种在定容条件下对生物体进行低温保存的方法。一种方法是isochoric freezing(译作“定容冷冻”或“等容冷冻”),通过利用等容腔体内部分液体发生凝固,产生的冰膨胀所带来的压力来降低剩余液体的凝固点温度,而生物材料以过冷态保存在剩余液体内,不存在冰晶损伤。另一种方法是定容过冷保存,通过利用整个等容腔体内部系统处于过冷态,从而使生物材料处于过冷态也不存在冰晶损伤。主要对定容冷冻和定容过冷两种保存方法的原理、应用和研究进展进行综述,并展望了定容低温保存技术未来的研究方向。
关键词： 冷冻保存;  过冷保存;  定容冷冻;  定容过冷;  冰物理    
Abstract:

Low temperature can slow down biochemical reaction and extend the life of biological materials. In order to avoid the cryoinjury caused by the ice crystal during traditional isobaric (atmospheric pressure) freezing process, Dr. Rubinsky developed two new biopreservation technologies at isochoric (constant volume) conditions. One technology is isochoric freezing, during which part of liquid is frozen and the formed ice expands to generate hydrostatic pressure inside the rigid isochoric chamber and biomatters can be stored at subfreezing temperatures in supercooled phase without any internal ice formation damage. The other technology is isochoric supercooling, which can also preserve organisms in supercooling state with a higher stability in a rigid isochoric chamber, resulting in no damage from ice crystals. In this paper, the principle, application and research progress of isochoric freezing and isochoric supercooling are reviewed, and the possible future research direction of cryopreservation under isochoric conditions is prospected.
Key words: Cryopreservation    Supercooling    Isochoric freezing    Isochoric supercooling    Ice physics
收稿日期: 2021-08-26 出版日期: 2022-03-03
ZTFLH:  Q-1  
基金资助: * 国家重点研发计划(2018YFD0400605);北京市科技计划(Z1711000013 17016);中国科学院青年创新促进会基金(2019030)
通讯作者: 陈六彪     E-mail: chenliubiao@mail.ipc.ac.cn
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引用本文:

赵远恒,郭嘉,陈六彪,王俊杰. 定容条件下生物低温保存研究进展*[J]. 中国生物工程杂志, 2022, 42(1/2): 119-127.

ZHAO Yuan-heng,GUO Jia,CHEN Liu-biao,WANG Jun-jie. A Review on Bio-preservation at Sub-freezing Temperatures under Isochoric Conditions. China Biotechnology, 2022, 42(1/2): 119-127.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2108059    或    https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I1/2/119

图1  常压冷冻、超高压冷冻、定容冷冻和过冷保存对比
图2  定容冷冻腔体[9]
图3  定容腔体中的典型冷冻过程
图4  定容冷冻保存鱼体组织[28,29]
图5  定容冷冻系统中添加保护液
生物材料 温度/℃ 保存效果 参考文献
小鼠肺 -2 结构损坏较小,大多数指标和新鲜肺几乎一致 Okamoto等[45]
小鼠肾脏 -2 组织结构破坏小 Sultana等[46]
人体肝脏 -4 有效延长肝脏活性至27 h de Vries等[47]
红细胞 -20 有效保存100天 Huang等[10]
表1  几种常见生物样品过冷保存结果
图6  不同条件下过冷液体稳定性:从固定高度撞击扰动后[45]
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