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临床输血与检验

临床输血与检验 ›› 2018, Vol. 20 ›› Issue (2): 141-144.DOI: 10.3969/j.issn.1671-2587.2018.02.010

• 临床输血 • 上一篇    下一篇

冰冻-解冻-去甘油过程红细胞形态变化与损伤相关性的探讨

林豪, 黄文华, 江伟梅, 林建霞, 陈灯锦   

  1. 350004 福州,福建省血液中心血液制备科
  • 收稿日期:2017-09-28 出版日期:2018-04-20 发布日期:2018-04-25
  • 作者简介:林豪（1974-）,男,福建福州人,副主任技师,学士,主要从事血液制备工作,（Tel）13959180746（E-mail）1900690083@qq.com。

The Study on the Correlation Between the Changes of RBCs Morphology and Injury in the Process of Freezing- thawing-Deglycerolizing

LIN Hao, HUANG Wen-hua, JIANG Wei-mei, et al   

  1. Fujian Blood Center 350004
  • Received:2017-09-28 Online:2018-04-20 Published:2018-04-25

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摘要： 目的 探讨冰冻-解冻-去甘油过程中红细胞形态变化与其损伤的相关性。 方法 依据配对设计原理,采用冻存前保留红细胞外多余甘油溶液（方法1）和冻存前离心去除红细胞外多余甘油溶液（方法2）的两种常规方法,制备冰冻解冻去甘油红细胞,对冰冻-解冻-去甘油过程各关键制备节点留取的样本,进行血常规及上清游离血红蛋白(FHb)含量的检测,分析红细胞形态变化及其与损伤的关联。 结果 ①冰冻-解冻-去甘油过程红细胞因甘油化而体积增大,后因去甘油化而复原,方法2较之方法1甘油化红细胞（冻存前）和冰冻解冻去甘油红细胞的平均体积（MCV）增高（P<0.01,P<0.05）;②冰冻-解冻-去甘油过程中红细胞损伤分布依次为:去甘油化过程(63.20±10.99) %、甘油化过程（30.78±10.47）%、冰冻解冻过程（6.02±4.11）%,两种制备方法间差异无统计学意义;③红细胞破坏与甘油化过程红细胞体积变化率呈中度负相关;④随着贮存时间的延长,冰冻解冻去甘油红细胞上清游离血红蛋白含量逐渐增加,0.9 %氯化钠悬浮冰冻解冻去甘油红细胞储存12 h、24 h,MAP悬浮冰冻解冻去甘油红细胞储存21 d、28 d,两种制备方法间差异有统计学意义（P<0.05）。 结论 冰冻-解冻-去甘油过程中红细胞损伤与其变形能力呈负相关,且损伤主要发生在去甘油化过程,冻存前离心去除红细胞外多余甘油溶液导致红细胞膜骨架因剪切力作用而受损,进而对冰冻解冻去甘油红细胞贮存质量产生不利影响。

关键词: 冰冻, 解冻, 去甘油, 红细胞, 平均红细胞容积, 损伤

Abstract: Objective To study the correlation between the changes of RBCs morphology and injury in the process of freezing-thawing-deglycerolizing. Methods The matching design method was employed to prepare frozen RBCs and thawed deglycerolized RBCs. Method1 : without supernatant glycerol reduction before being frozen. Method2 : supernatant glycerol reduction by centrifugation before being frozen. Take samples for blood routine test and free hemoglobin (FHb) content detection in the process of freezing-thawing-deglycerolizing, analysis the shape variation of red blood cells and its correlation with injury. Result (1) In the process of freezing-thawing-deglycerolizing, volume of red blood cell increase due to glycerolize ,and then recovery due to deglycerolize. Compare with the method 2 and method 1, mean corpuscular volume (MCV) of glycerolized RBCs (before being frozen) and thawed deglycerolized RBCs increased (P<0.01, P<0.05);(2) The destruction of red blood cells is distributed in three processes, deglycerolizing process (63.20±10.99) %, glycerolizing process (30.78±10.47) %, freezing-thawing process (6.02±4.11) %, no statistical difference between two kinds of preparation methods.(3) the red blood cell damage is moderate negative correlation with volume change rate of glycerolized RBCs ; (4) with the extension of storage time, free hemoglobin content of thawed deglycerolized RBCs gradually increased. In which the 0.9% NaCl suspension thawed deglycerolized RBCs, the content of FHb showed significantly different 12 h and 24 h after preparation as well (P<0.05).In which the MAP suspension thawed deglycerolized RBCs, the content of FHb showed significantly different 21d and 28d after preparation as well (P<0.05). Conclusion In the process of freezing-thawing-deglycerolizing, red blood cell damage is negatively related to its deformation ability, and the damage mainly happens on the deglycerolizing process. The removal of supernatant glycerol by centrifugation caused the red cell membrane skeleton to be damaged by the shear stress, thereby adversely affecting the storage quality of the thawed deglycerolized RBCs.

Key words: Freezing, Thawing, Deglycerolizing, Red blood cells, MCV, Injury

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