红细胞冻干保存中的渗透性损伤实验研究

红细胞长期保存中保护剂添加、洗涤过程会引入红细胞渗透性损伤.在冻干保存研究中,由于多种保护剂同时使用,保护剂的类型和功能一直是研究的重点,但很少有从渗透性损伤角度分析保存方案合理性的报道.目前相关文献、专利中所用的保护剂总渗透压差别很大,细胞保存后回收率差异也较大.文中用NaCl溶液实验模拟红细胞保存中保护剂添加、洗涤过程.结果表明,选择合适的添加、洗涤方法可以在一定程度上减小渗透损伤.就红细胞而言,1.5Osmol/kg左右是保护剂总渗透压的一个重要阈值,总渗透压低于该阈值时,渗透性损伤较小;高于该阈值时,渗透损伤随着渗透压的增大而迅速增大.所以选择保护剂时,首先应该根据总渗透压来排除渗透压过高的保存方案,否则红细胞在添加和洗涤保护剂时已经损伤很大.该研究对其它细胞长期保存中保护剂的选择也具有参考意义.

The Experimental Study of Osmotic Injury to Erythrocytes in Freeze Drying

In the long-term preservation of red blood cells,the osmotic injury occurs during the addition and removal of the protective agents,which is often omitted in practice.In the freeze drying,many kinds of agents are used together.Now the research commonly focuses on the type and function of the agents.But there are still few reports of the analysis on the rationality of the preservation protocols with the view of osmotic injury.In the present relevant papers and patents,the osmolalities of the protective agents varied greatly.The experiment results also differed greatly.It was argued in this paper that the good agents could only work well with optimum osmolality.Different concentrations of NaCl solutions were used to simulate the addition and removal of protective agents.The results showed that the osmotic injury could be decreased with optimized addition and removal procedures.For erythrocytes,the critical osmolality of the agents was found to be about 1.5Osmol/kg.Above that,the cells would be lost greatly just during the addition and removal procedure.It was demonstrated that it was important for one to check the osmolality of the protective agents firstly in the erythrocyte preservation,especially in freeze drying.This work is also helpful for the long-term preservation of other cells.