Ameri Zahra, Farsinejad Alireza, Vahidi Reza, Sheikh Rezaei Zahra, Khaleghi Morteza, Parvizi Poorya, Moghadari Masoud
Student Research Committee, Kerman University of Medical Sciences, Kerman, Iran.
Department of Tissue Engineering and Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
Indian J Hematol Blood Transfus. 2022 Apr;38(2):373-380. doi: 10.1007/s12288-021-01447-4. Epub 2021 May 17.
The present study aims to investigate the changes in different parameters related to the storage time of red blood cell (RBC) units. Microscopic, flow cytometric, and electrophoretic assessments were employed every few days for 60 days to investigate the alterations in morphology, size, phosphatidylserine (PS) externalization, and membrane proteins over time. Morphological transformation from discocytes to spherocytes progressed as the storage time increased, which was accompanied by an increment of cellular size. However, this storage period did not result in the externalization of significant amounts of PS ( > 0.05). Mean Fluorescence Intensity (MFI) values increased by 11% to 23% between days 21 and 35 compared to the day 1 sample ( < 0.001). By day 60, the MFI decreased to about 70% of the day 1 sample. The analysis of membrane proteins' distribution showed a significant drop in band 3 expression after 35 days ( < 0.05 and 0.001 on days 42 and 60, respectively); however, no significant change was observed up to five weeks ( > 0.05). The inconsistency observed between Eosin-5-Maleimide (5-EMA) binding and the relative band 3 content could be due to additional accessibility of 5-EMA to hidden domains of other membrane proteins on RBCs as a result of increased mean corpuscular volume (MCV) and changes in morphology. Overall, our present study represents a step-wise and time-dependent series of events that progressively affects stored RBCs.
本研究旨在调查与红细胞（RBC）单位储存时间相关的不同参数的变化。每隔几天进行一次为期60天的显微镜、流式细胞术和电泳评估，以研究形态、大小、磷脂酰丝氨酸（PS）外化和膜蛋白随时间的变化。随着储存时间的增加，红细胞从双凹圆盘状向球形的形态转变逐渐进展，同时细胞大小增加。然而，这个储存期并未导致大量PS的外化（>0.05）。与第1天的样本相比，第21天至35天期间平均荧光强度（MFI）值增加了11%至23%（<0.001）。到第60天，MFI降至第1天样本的约70%。膜蛋白分布分析显示，35天后带3表达显著下降（第42天和第60天分别为<0.05和<0.001）；然而，在五周内未观察到显著变化（>0.05）。观察到的嗜酸性-5-马来酰亚胺（5-EMA）结合与相对带3含量之间的不一致，可能是由于平均红细胞体积（MCV）增加和形态变化导致5-EMA对红细胞上其他膜蛋白隐藏结构域的额外可及性。总体而言，我们目前的研究代表了一系列逐步的、随时间变化的事件，这些事件逐渐影响储存的红细胞。
