Storage-Induced Micro-Erythrocytes Can Be Quantified and Sorted by Flow Cytometry.
Storage-Induced Micro-Erythrocytes Can Be Quantified and Sorted by Flow Cytometry.
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DOI：
10.3389/fphys.2022.838138
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发表时间：
2022
影响因子：
4
通讯作者：
Amireault P
中科院分区：
文献类型：
--
作者：
Marin M;Peltier S;Hadjou Y;Georgeault S;Dussiot M;Roussel C;Hermine O;Roingeard P;Buffet PA;Amireault P
输血前冷藏红细胞浓缩物与红细胞（RBC）的进行性改变有关。小RBC（III型棘红细胞、球形棘红细胞和球形红细胞）定义为储存诱导的微小红细胞（SME），在输血前储存期间出现。SME在不同供者间的积累强度不同，在输血后迅速清除，其比例与输血回收率相关。它们可以使用成像流式细胞术（IFC）快速客观地定量。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 ℃下孵育。通过流式细胞术评估CFSE强度，并通过IFC评估RBC形态。我们通过流式细胞术观察到CFSE高RBC亚群的积累，在储存的第3天和第42天分别占3.3%和47.2%。IFC明视野图像显示，CFSE高亚群主要包含SME，而CFSE低亚群主要包含I型和II型棘细胞和椎间盘细胞。通过IFC（基于投影表面积）和流式细胞术（基于CFSE强度）定量相似数量的SME。IFC和扫描电镜显示，通过基于流式细胞术的分选获得了纯度≥95%的CFSE high和CFSE low 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。染色方案实现了SME的特异性分选，这是进一步表征输血后过早清除的RBC亚群的有用工具。
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 CFSEhigh 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 CFSEhigh subpopulation mostly contains SME while the CFSElow 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 CFSEhigh and CFSElow 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.
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影响因子：
2.9
作者：
Lu, Madeleine;Shevkoplyas, Sergey S.
通讯作者：
Shevkoplyas, Sergey S.
影响因子：
3.7
作者：
Banks, H. T.;Choi, A.;Thompson, W. C.
通讯作者：
Thompson, W. C.
影响因子：
6.5
作者：
Burger, Patrick;Kostova, Elena;van Bruggen, Robin
通讯作者：
van Bruggen, Robin
影响因子：
3.7
作者：
Bardyn, Manon;Rappaz, Benjamin;Prudent, Michel
通讯作者：
Prudent, Michel
DOI：
10.1186/cc12600
发表时间：
2013-04-08
期刊：
Critical care (London, England)
影响因子：
--
作者：
Lelubre C;Vincent JL
通讯作者：
Vincent JL