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Theoretical and experimental analyses of optimal experimental design for determination of hydraulic conductivity of cell membrane

机译:测定细胞膜水导率的最佳实验设计的理论和实验分析

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摘要

Determination of cell hydraulic conductivity (L_p) is required to predict the optimal conditions for cell cryopreservation. One of the critical procedures associated with the determination of L_p is to measure the kinetics of cell volume change in response to a sudden cell exposure to anisosmotic media until the cells achieve an osmotic equilibrium state. To achieve accurate measurement, it should be ensured that (1) the cell osmotic equilibration process is sufficiently slow, and (2) the total cell volume change (ΔV) is much larger than the resolution of the measuring device (δ). In this article, a cell's half volume excursion time (t~*) was defined as the time in which osmotically active cell water volume increases or decreases by half of its maximum change. Based on the water transport equations, a series of analytical solutions were derived. The t~* and ΔV were expressed as functions of 2 control variables: initial intracellular osmolality (M_o) and extracellular osmolality (M_e), and the effects of M_e and M_o on t~* and ΔV were predicted theoretically. The predictions were confirmed by performing experiments using two different cell types. In the light of this study, a strategy to optimize the experiment design for the L_p determination is suggested.
机译:确定细胞的水力传导率(L_p)是预测细胞冷冻保存的最佳条件所必需的。与确定L_p相关的关键程序之一是测量细胞突然改变接触异渗介质的反应,直至细胞达到渗透平衡状态,从而测量细胞体积变化的动力学。为了实现准确的测量,应确保(1)细胞渗透平衡过程足够缓慢,以及(2)总细胞体积变化(ΔV)远大于测量设备的分辨率(δ)。在本文中,细胞半体积偏移时间(t〜*)定义为渗透活性细胞水体积增加或减少最大变化一半的时间。根据输水方程,得出了一系列的解析解。将t〜*和ΔV表示为两个控制变量的函数:初始细胞内渗透压(M_o)和细胞外渗透压(M_e),并从理论上预测了M_e和M_o对t〜*和ΔV的影响。通过使用两种不同的细胞类型进行实验,证实了这一预测。根据这项研究,提出了一种针对L_p测定优化实验设计的策略。

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