Kinetics and mechanism of intercellular ice propagation in a micropatterned tissue construct
Kinetics and mechanism of intercellular ice propagation in a micropatterned tissue construct
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DOI：
10.1016/s0006-3495(02)75536-7
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发表时间：
2002-04-01
影响因子：
3.4
通讯作者：
Karlsson, JOM
中科院分区：
文献类型：
--
作者：
Irimia, D;Karlsson, JOM
了解细胞间相互作用对细胞内冰形成（IIF）的影响是设计组织冷冻保存的优化方案所必需的。为了确定组织冷冻过程中细胞-细胞相互作用的影响，而不受不受控制的因素（如培养时间，细胞几何形状和细胞-基质相互作用）的混杂影响，将HepG 2细胞成对培养在用聚乙二醇二硅烷微图案化的玻璃盖玻片上，使得每个细胞恰好与一个相邻细胞相互作用。假设细胞对是一个有限状态系统，处于未冻结状态（任一细胞中没有冰），单线态（仅一个细胞中的IIF）或双线态（两个细胞中的IIF），状态转变的动力学理论建模和低温显微镜测量。从测得的单态概率估计的细胞间冰的繁殖率，在培养的第一个24小时增加，此后保持稳定。在培养24小时并在冷冻前用差距连接阻断剂18 β-甘草次酸处理的细胞对中，细胞间冰繁殖率低于未处理的对照组（p < 0.001），但显着大于零（rho < 0.0001）。这些结果表明，间隙连接介导的一些，但不是全部，冰在组织中传播的机制。
Understanding the effects of cell-cell interaction on intracellular ice formation (IIF) is required to design optimized protocols for cryopreservation of tissue. To determine the effects of cell-cell interactions during tissue freezing, without confounding effects from uncontrolled factors (such as time in culture, cell geometry, and cell-substrate interactions), HepG2 cells were cultured in pairs on glass coverslips micropatterned with polyethylene glycol disilane, such that each cell interacted with exactly one adjacent cell. Assuming the cell pair to be a finite state system, being either in an unfrozen state (no ice in either cell), a singlet state (IIF in one cell only), or a doublet state (IIF in both cells), the kinetics of state transitions were theoretically modeled and cryomicroscopically measured. The rate of intercellular ice propagation, estimated from the measured singlet state probability, increased in the first 24 h of culture and remained steady thereafter. In cell pairs cultured for 24 h and treated with the gap junction blocker 18beta-glycyrrhetinic acid before freezing, the intercellular ice propagation rate was lower than in untreated controls (p < 0.001), but significantly greater than zero (rho < 0.0001). These results suggest that gap junctions mediate some, but not all, mechanisms of ice propagation in tissue.