PDF(Pubmed)
Abstract：
Refrigerated storage of red cell concentrates before transfusion is associated with progressive alterations of red blood cells (RBC). Small RBC (type III echinocytes, sphero-echinocytes, and spherocytes) defined as storage-induced micro-erythrocytes (SME) appear during pretransfusion storage. SME accumulate with variable intensity from donor to donor, are cleared rapidly after transfusion, and their proportion correlates with transfusion recovery. They can be rapidly and objectively quantified using imaging flow cytometry (IFC). Quantifying SME using flow cytometry would further facilitate a physiologically relevant quality control of red cell concentrates. RBC stored in blood bank conditions were stained with a carboxyfluorescein succinimidyl ester (CFSE) dye and incubated at 37°C. CFSE intensity was assessed by flow cytometry and RBC morphology evaluated by IFC. We observed the accumulation of a CFSE high RBC subpopulation by flow cytometry that accounted for 3.3 and 47.2% at day 3 and 42 of storage, respectively. IFC brightfield images showed that this CFSE high subpopulation mostly contains SME while the CFSE low subpopulation mostly contains type I and II echinocytes and discocytes. Similar numbers of SME were quantified by IFC (based on projected surface area) and by flow cytometry (based on CFSE intensity). IFC and scanning electron microscopy showed that ≥95% pure subpopulations of CFSE high and CFSE low RBC were obtained by flow cytometry-based sorting. SME can now be quantified using a common fluorescent dye and a standard flow cytometer. The staining protocol enables specific sorting of SME, a useful tool to further characterize this RBC subpopulation targeted for premature clearance after transfusion.
摘要：
输血前红细胞浓缩物的冷藏与红细胞（RBC）的进行性改变有关。小红细胞（III型棘突细胞，棘球细胞,和球形细胞）定义为在输血前储存期间出现的储存诱导的微红细胞（SME）。中小企业以不同的强度从供体到供体积累，输血后迅速清除，它们的比例与输血恢复有关。可以使用成像流式细胞术（IFC）快速客观地定量它们。使用流式细胞术定量SME将进一步促进红细胞浓缩物的生理学相关质量控制。将储存在血库条件下的RBC用羧基荧光素琥珀酰亚胺酯(CFSE)染料染色并在37°C下孵育。通过流式细胞术评估CFSE强度,通过IFC评估RBC形态。我们通过流式细胞术观察到CFSE高红细胞亚群的积累，在储存的第3天和第42天分别占3.3％和47.2％，分别。IFC明场图像显示，该CFSE高亚群主要包含SME，而CFSE低亚群主要包含I型和II型棘突细胞和椎间盘细胞。通过IFC(基于投影表面积)和通过流式细胞术(基于CFSE强度)定量类似数量的SME。IFC和扫描电子显微镜显示，通过基于流式细胞术的分选获得了≥95％纯的CFSE高和CFSE低RBC亚群。现在可以使用普通荧光染料和标准流式细胞仪定量SME。染色协议可以对SME进行特定的分类，一个有用的工具，以进一步表征这种针对输血后过早清除的RBC亚群。
