冷冻保存中影响胞内冰生长的因素

doi:10.11949/0438-1157.20190552

摘要/Abstract

摘要：

细胞胞内冰的形成会导致严重的细胞损伤从而导致低温贮存中的诸多问题。以蚕豆为研究对象，用细胞松弛素B溶解细胞骨架，使用低温显微系统在不同的冷却速率下进行冷冻实验。实验结果表明，使用细胞松弛素B处理过的细胞在冷冻过程中结晶温度更高，结晶时间更短，但细胞骨架对胞内冰的生长过程影响较小。外界条件起着关键作用，接种冰晶影响细胞内冰晶的形成温度及冰晶的生长速率。最后，通过光强度图对细胞的损伤程度进行了分析。

关键词: 显微测量, 微丝骨架, 细胞松弛素B, 传质, 细胞生物学, 界面

Abstract:

The formation of intracellular ice of cells can cause severe cellular damage leading to many problems in cryopreservation. The broad bean was used as the research object, and the cytoskeleton was dissolved by cytochalasin B, and the freezing experiment was carried out at different cooling rates using a low temperature microscopic system. The experimental results show that the cells treated with cytochalasin B have higher crystallization temperature and shorter crystallization time during the freezing process, but the cytoskeleton has little effect on the growth process of intracellular ice. The external conditions play a key role, and the inoculation of ice crystals affects the formation temperature of ice crystals in the cells and the growth rate of ice crystals. Finally, the degree of damage to the cells was analyzed by light intensity map. It was found that the growth process of intracellular ice without cytoskeleton was similar to that of normal cells, but the crystallization rate was different, indicating that the cytoskeleton inside the cells affected the initiation of intracellular ice. Therefore, keeping the cytoskeleton intact before cryopreserving the plant sample can reduce the formation of intracellular ice and maintain cell morphology after rewarming.

Key words: microscopic measurement, microfilament skeleton, cytochalasin B, mass transfer, cell biology, interface

中图分类号:

TB 61

诸凯, 谢艳琦, 王雅博. 冷冻保存中影响胞内冰生长的因素[J]. 化工学报, 2019, 70(S2): 208-214.

Kai ZHU, Yanqi XIE, Yabo WANG. Factors affecting intracellular ice growth during cryopreservation[J]. CIESC Journal, 2019, 70(S2): 208-214.

图/表 11

图1 正常细胞中冰晶体生长的过程

Fig.1 Process of ice crystal growth in normal cells

图2 没有细胞骨架的细胞中冰晶体生长的过程

Fig.2 Process of ice crystal growth in cells without cytoskeleton

图3 不同细胞中冰晶生长行为出现的比例

Fig.3 Proportion of ice crystal growth behavior in different cells

图4 没有接种冰晶的细胞内冰晶生长速率

Fig.4 Ice crystal growth rate of cells not inoculated with ice crystals

图5 接种冰晶后细胞内冰晶生长速率

Fig.5 Ice crystal growth rate of cells inoculated with ice crystals

图6 没有接种冰晶的细胞

Fig.6 Cells not inoculated with ice crystals

图7 接种冰晶的细胞

Fig.7 Cells inoculated with ice crystals

图8 室温下细胞的形态

Fig.8 Cell morphology at room temperature

图9 冷冻之前的3D-光强图

Fig.9 3D-light intensity map before freezing

图10 复温到25℃的3D-光强图

Fig.10 3D-light intensity map rewarmed to 25 °C

图11 不同冷却速率下的光强度

Fig.11 Light intensity at different cooling rates

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