Alshalani Abdulrahman, Acker Jason P
Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada; Department of Clinical Laboratory Sciences, King Saud University, Riyadh, Saudi Arabia.
Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada; Centre for Innovation, Canadian Blood Services, Edmonton, Alberta, Canada.
Cryobiology. 2017 Jun;76:51-58. doi: 10.1016/j.cryobiol.2017.04.009. Epub 2017 Apr 26.
Water transport across the red blood cell (RBC) membrane is an essential cell function that needs to be preserved during ex vivo storage. Progressive biochemical depletion during storage can result in significant conformational and compositional changes to the membrane. Characterizing the changes to RBC water permeability can help in evaluating the quality of stored blood products and aid in the development of improved methods for the cryopreservation of red blood cells. This study aimed to characterize the water permeability (L), osmotically inactive fraction (b), and Arrhenius activation energy (E) at defined storage time-points throughout storage and to correlate the observed results with other in vitro RBC quality parameters. RBCs were collected from age- and sex-matched blood donors. A stopped flow spectrophotometer was used to determine L and b by monitoring changes in hemoglobin autofluorescence when RBCs were exposed to anisotonic solutions. Experimental values of L were characterized at three different temperatures (4, 20 and 37 °C) to determine the E. Results showed that L, b, and E of stored RBCs significantly increase by day 21 of storage. Degradation of the RBC membrane with length of storage was seen as an increase in hemolysis and supernatant potassium, and a decrease in deformability, mean corpuscular hemoglobin concentration and supernatant sodium. RBC osmotic characteristics were shown to change with storage and correlate with changes in RBC membrane quality metrics. Monitoring water parameters is a predictor of membrane damage and loss of membrane integrity in ex vivo stored RBCs.
水通过红细胞（RBC）膜的转运是一项基本的细胞功能，在体外储存过程中需要保持。储存期间的渐进性生化消耗会导致膜发生显著的构象和成分变化。表征红细胞水渗透性的变化有助于评估储存血液制品的质量，并有助于开发改进的红细胞冷冻保存方法。本研究旨在表征在整个储存过程中特定储存时间点的水渗透性（L）、渗透非活性部分（b）和阿伦尼乌斯活化能（E），并将观察结果与其他体外红细胞质量参数相关联。红细胞取自年龄和性别匹配的献血者。使用停流分光光度计通过监测红细胞暴露于非等渗溶液时血红蛋白自发荧光的变化来测定L和b。在三个不同温度（4、20和37°C）下对L的实验值进行表征以确定E。结果表明，储存红细胞的L、b和E在储存第21天时显著增加。随着储存时间的延长，红细胞膜的降解表现为溶血和上清钾增加，以及变形性、平均红细胞血红蛋白浓度和上清钠降低。红细胞渗透特性显示随储存而变化，并与红细胞膜质量指标的变化相关。监测水参数是体外储存红细胞膜损伤和膜完整性丧失的一个预测指标。
