再水化液因素对冰冻干燥保存后红细胞回收率的影响

目的　寻求一种能有效提高冰冻干燥 (简称冻干 )保存后人红细胞回收率的再水化体系。方法　测定1 0 %聚乙烯吡咯烷酮 (PVP)、6 %羟乙淀粉 (HES)、5 %羧甲基淀粉钠 (CMS)、生理盐水、0 75mol/L葡萄糖、等渗缓冲液及高渗缓冲液 (5×Buffer)的晶体渗透压和胶体渗透压 ;将浓缩红细胞和保护液混匀 ,预冻后移入冻干机内作冻干处理 ,冻干完毕后 ,用不同种类或不同温度的再水化液快速水化洗涤样本。结果　人红细胞冻干再水化后 ,6 %HES组、1 0 %PVP组和 5 %CMS组的红细胞回收率分别为 (93.6 5± 6 .1 8) %、(88.80± 9.4 9) %和 (91 .34± 8.1 3) % ,血红蛋白回收率分别为 (93.4 8± 4 .6 7) %、(89.0 2± 4 .6 7) %和 (88.79± 5 .35 ) % ,均极显著高于其他 4组[(1 5 .5 6± 1 2 .0 2 ) %～ (2 7.77± 6 .4 8) % ,(1 7.78± 1 0 .80 ) %～ (4 1 .5 0± 6 .4 3) % ) ](P <0 .0 1 ) ;不同温度的 6 %HES的再水化效果表明 ,再水化后 3个温度组的红细胞回收率无显著差异 ,但 37℃和 2 5℃组的血红蛋白回收率分别为(87.4 8± 5 .84 ) %和 (91 .37± 3.94 ) % ,均极显著高于 4℃组 (73.1 0± 5 .90 ) % (P <0 .0 1 )而且上清游离血红蛋白浓度也显著低于 4℃组。结论　再水化液的胶体渗透压对冻干保存后红细胞的保护作

Effect of reconstitution solutions on the recovery of lyophilized red blood cells

Objectives To maximize the survival of lyophilized red blood cells after reconstitution.Methods The colloidal and crystalloid osmotic pressures of seven reconstitution solutions,including 10% polyvinylpyrrolidone(PVP),6% hydroxyethyl starch (HES),5% carboxymethylcellulose sodium (CMS),0.9%NaCl,0.75mol/L Glucose,isotonic buffer ,and hypertonic buffer (5×buffer),were measured. After washing and equilibrating,red blood cells were mixed with protective solution according to the ratio of 1∶3,then pre frozen at -80℃ for 1h,at last,samples were lyophilized in freeze dryer.After lyophilization,the samples were reconstituted in different reconstitution solutions and at different temperatures,the red blood cell recovery was measured.Results After lyophilization and reconstitution,the recovery of red blood cells in 6%HES,10%PVP,and 5%CMS were (93.65±6.18)%,(88.80±9.49)% and (91.34±8.13)%,respectively,and the recovery of hemoglobin were (93.48±4.67)%,(89.02±4.67)%,and (88.79±)5.35% respectively,significantly higher than those of the other groups[(15.56±12.02)%～(27.77±6.48)%,(17.78±10.80)%～(41.50±6.43)%] (P <0.01). At temperatures of 4℃,25℃,and 37℃,the recovery of red blood cells were not different by using 6%HES as reconstitution solution. But the recovery of hemoglobin at 37℃ and 25℃ were 87.48±5.84% and 91.37±3.94%,significantly higher than at 4℃ (73.10±5.90%) ( P <0.01).The concentrations of free hemoglobin at 25℃ and 37℃ were significantly less than at 4℃.Conclusion The effect of colloidal osmotic pressure on the survival of red blood cells after lyophilization and reconstitution is better than that of crystalloid,so addition of low concentration polymers in isotonic buffer as reconstitution solutions is beneficial for survival of red blood cells after lyophilization. The recovery is better at temperatures higher than 25℃,compared to at 4℃.