人红细胞冻干后复水条件的初步研究

Preliminary Study on Rehydrated Conditions for Lyophilized Human Red Blood Cells

本研究旨在探讨不同复水条件对冻干红细胞回收率的影响,为冻干红细胞复水条件的优化提供实验依据。在一系列不同条件下,包括复水溶液、复水温度、复水阶段红细胞胞体积变化速率(即蒸汽预复水)等,对冻干红细胞进行复水,检测红细胞复水后回收率并检测冻干红细胞复水后各项理化功能的改变。结果显示:在复水液配方中以PBS为基液配制的10%(w/v)PVP40溶液效果较好。在复水温度方面,低于37℃时红细胞回收率随着温度的升高而增加,42℃比37℃略差,37℃下复水效果最好。在加入复水液之前将样品放置在37℃的饱和蒸汽环境中预复水能提高红细胞回收率并使细胞体积和体积分布宽度更接近于新鲜细胞数值。复水后红细胞理化性质的检测显示,红细胞的变形性较常规保存的红细胞有所下降,但胞内ATP酶、G-6-PD酶、SOD和2,3-DPG的活性降低较少,酶活性回收率大于红细胞回收率。结论:人红细胞冻干后最佳的复水条件是:冻干红细胞在37℃的饱和蒸汽环境中预复水,复水液为PBS+10%PVP40,复水温度为37℃,但对冻干复水过程中的细胞膜的保护还需进行更深入的研究。

The objective of this study was to investigate the effect of different rehydration conditions on recovery of the lyophilized red blood cells （RBC） so as to optimize the RBC rehydration. The different conditions, including diffe- rent rehydration solution, the rehydration temperature, volume change rate of the lyophilized RBC rehydrated by the vapor firstly, were studied, the recovery rate and change of physiological and biochemical properties of the rehydrated RBC were detected. The results indicated that the solution of 10% （w/v）PVP40 in PBS showed the best effect, and the RBC recovery rate increased with increasing of rehydration temperature, and the optimal temperature of rehydration was at 37℃. Pre-rehydration in condition of vapor could raise the RBC recovery rate, and promote the MCV and RDW to close to index of the fresh RBC, the deformability of the rehydrated RBC was no serious as compared with RBC preserved in conventional condition, but the activity level of ATP, G-6-PD, SOD, 2,3-DPG of the rehydrated RBC less decreased. It is concluded that the optimal rehydration conditions for lyophilized RBC are pre-rehydration in the 37℃ with vapor firstly, PBS ＋ 10% （w/v） PVP40 rehydration solution and rehydration temperature at 37℃, but the protection of RBC membrane needs to be furtherly studied.