Cell-cell contact affects membrane integrity after intracellular freezing
Cell-cell contact affects membrane integrity after intracellular freezing
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DOI：
10.1006/cryo.1999.2221
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发表时间：
2000-02-01
期刊：
影响因子：
2.7
通讯作者：
McGann, LE
中科院分区：
文献类型：
--
作者：
Acker, JP;McGann, LE
细胞对冷冻的反应主要取决于完整细胞膜的存在。在快速冷却过程中，细胞质膜可能不再是冰传播的有效屏障，并且可以被细胞外冰破坏，导致过冷细胞质的成核。在组织中，细胞内冰的形成是由细胞-细胞和细胞-表面相互作用的存在而复合的。使用三种不同的仓鼠成纤维细胞模型系统来模拟在组织化组织中发现的结构。样品在低温台上过冷到实验温度，并在恒定温度下成核。使用双重荧光染色技术定量评估细胞质膜的完整性。一种新的技术，使用荧光染料SYTO用于检测细胞单层的细胞内冰形成（IIF）。细胞膜完整性丧失的累积发生率和IIF的累积发生率被确定为温度的函数。悬浮液中的细胞和单个附着的细胞在形成细胞内冰的细胞数量和失去膜完整性的细胞数量方面没有显着差异。对于单层细胞，在细胞-细胞接触的情况下，细胞内冰的形成不会导致大多数细胞的质膜立即破裂。这就有可能最大限度地减少IIF造成的损伤，并在快速冷却过程中制定组织冷冻保护策略。(C)北京大学出版社.
The response of cells to freezing depends critically on the presence of an intact cell membrane. During rapid cooling, the cell plasma membrane may no longer be an effective barrier to ice propagation and can be breached by extracellular ice resulting in the nucleation of the supercooled cytoplasm. In tissues, the formation of intracellular ice is compounded by the presence of cell-cell and cell-surface interactions. Three different hamster fibroblast model systems were used to simulate structures found in organized tissues. Samples were supercooled to an experimental temperature on a cryostage and ice nucleated at the constant temperature. A dual fluorescent staining technique was used for the quantitative assessment of the integrity of the cell plasma membrane. A novel technique using the fluorescent stain SYTO was used for the detection of intracellular ice formation (IIF) in cell monolayers. The cumulative incidence of cells with a loss of membrane integrity and the cumulative incidence of IIF were determined as a function of temperature. Cells in suspension and individual attached cells showed no significant difference in the number of cells that formed intracellular ice and those that lost membrane integrity. For cells in a monolayer, with cell-cell contact, intracellular ice formation did not result in the immediate disruption of the plasma membrane in the majority of cells. This introduces the potential for minimizing damage due to IIF and for developing strategies for the cryoprotection of tissues during rapid cooling. (C) 2000 Academic Press.