Abidor I G, Li L H, Hui S W
Membrane Biophysics Laboratory, Roswell Park Cancer Institute, Buffalo, New York 14263.
Biophys J. 1994 Jul;67(1):418-26. doi: 10.1016/S0006-3495(94)80497-7.
Cell pellets formed by centrifugation provided a good system to study the osmotic behavior, electroporation, and interaction between cells. Rabbit erythrocyte pellets were used in this study because they were simpler than nucleated cells to model analytically. Structurally, cell pellets possessed properties of porous solid bodies and gels. Electrically, cell pellets were shown to behave as a parallel set of resistance, Rp, and capacitance, Cp. Information on pellet structures was obtained from electric measurements. The pellet resistance reflected the intercellular conductivity (porosity and gap conductivity), whereas the pellet capacitance depended mostly on membrane capacitance. The pellet resistance was more sensitive to experimental conditions. The intercellular gap distance can be derived from pellet porosity measurements, providing the cell volume and surface area were known. Rp increased and relaxed exponentially with time when centrifugation started and stopped; the cycles were reversible. When supernatants were exchanged with solutions containing hypotonic electrolytes or macromolecules (such as PEG) after the pellets were formed, complicated responses to different colloidal osmotic effects were observed. A transient decrease followed by a large increase of Rp was observed after the application of a porating electric pulse, as expected from a momentary membrane breakdown, followed by a limited colloidal-osmotic swelling of pelleted cells. The equilibrium values of Rp, Cp, pellet porosity, and intercellular distances were measured and calculated as functions of cell number, centrifugation force, and ionic strength of the exchanged supernatant. Thus, the structure and properties of cell pellets can be completely characterized by electrical measurements.
通过离心形成的细胞沉淀提供了一个研究渗透行为、电穿孔以及细胞间相互作用的良好系统。本研究使用兔红细胞沉淀,因为它们比有核细胞在分析建模上更简单。从结构上看,细胞沉淀具有多孔固体和凝胶的特性。从电学角度看,细胞沉淀表现为一组并联的电阻(Rp)和电容(Cp)。通过电学测量获得了沉淀结构的信息。沉淀电阻反映了细胞间的电导率(孔隙率和间隙电导率),而沉淀电容主要取决于膜电容。沉淀电阻对实验条件更敏感。当已知细胞体积和表面积时,细胞间间隙距离可从沉淀孔隙率测量中得出。离心开始和停止时,(Rp)随时间呈指数增加和松弛;这些循环是可逆的。沉淀形成后,当用含有低渗电解质或大分子(如聚乙二醇)的溶液替换上清液时,观察到对不同胶体渗透效应的复杂反应。施加穿孔电脉冲后,如瞬间膜破裂所预期的那样,观察到(Rp)先短暂下降然后大幅增加,随后沉淀细胞出现有限的胶体渗透肿胀。测量并计算了(Rp)、(Cp)、沉淀孔隙率和细胞间距离的平衡值,作为细胞数量、离心力和交换上清液离子强度的函数。因此,细胞沉淀的结构和性质可以通过电学测量完全表征。