冰温保存肾表皮细胞及其渗透特性的实验研究

摘要

In this study,experiments were performed to determine the permeability characteristics of kidney epidermis cells at various temperatures and different osmotic pressures using microscopy image analysis,including osmotically inactive volume,membrane permeability of water,and membrane permeability of CPA. The mean volume at isotonic solution was Viso=2472.58μm3;the history of cell volume change conformed to Boyle van’t Hoff relationship. The experimental data then were fitted to a straight line by the linear least square method and extrapolated by linear regression to determine osmotically inactive volume Vb=0.22Viso. Permeability response to hypertonic CPA solution was observed. Membrane permeability of water (Lp) and membrane permeability of CPA (Ps) and its activation energy(Ea) in dimethyl sulfoxide (DMSO),ethylene glycol(EG) and glycerinum (GLY) solution were determined by using two-parameter transport model;Based on the survival rate data generated in ice-temperature and normal temperature storage,the optimal preservation temperature for kidney epidermis cells was obtained. Ice-temperature storage has bright application prospect in cryo-preservation.%以肾表皮细胞为研究对象，利用显微镜图像分析方法，研究了其在不同温度、不同渗透压下的渗透特性，其中包括细胞的非渗透体积、细胞膜对水和低温保护剂的渗透系数等。结果表明，在等渗溶液中，细胞的平均等渗体积Viso=2472.58μm3；细胞体积随溶液渗透压的变化规律符合Boyle van’t Hoff 关系式，利用最小二乘法对实验数据进行线性拟合处理，得到非渗透体积 Vb=0.22Viso；观察细胞在高渗低温保护剂溶液中的渗透响应，利用两参数模型，得到了不同温度下在二甲基亚砜、乙二醇、甘油溶液中肾表皮细胞对水的渗透系数（Lp）、对低温保护剂的渗透系数（Ps）及活化能（Ea）；通过对细胞在冰温和常温下进行保存，测量细胞的存活率，得到了针对肾表皮细胞的最佳保存温度。基于上述结果，说明冰温保存细胞的方法在低温保存领域具有良好的应用前景。