Laboratoire de Recherche sur les Produits Sanguins, Transfusion Interrégionale CRS SA, Epalinges, Switzerland.
Faculté de Biologie et de Médecine, University of Lausanne (UNIL), Lausanne, Switzerland.
Blood Transfus. 2023 Jul;21(4):277-288. doi: 10.2450/2022.0086-22. Epub 2022 Nov 4.
The quality of red blood cells (RBCs) stored in red cell concentrates (RCCs) is influenced by processing, storage and donor characteristics, and can have a clinical impact on transfused patients. To evaluate RBC properties and their potential impact in a transfusion setting, a simple in vitro-transfusional model has been developed.
Transfusion was simulated by mixing a washed RBC pool from two male-derived RCCs stored at 4°C with a pool of 15 male-derived fresh frozen plasma (FFP) units, representing the recipient, at a hematocrit (HCT) of 30% ("control" setting) or 5% (alternative model). The mixtures were incubated at 37°C, 5% of CO up to 48 h. Different metabolites, hemolysis and microvesicles (MVs) were quantified at several incubation times and RBC-morphology changes and deformability after incubation. For each model, biological triplicates have been investigated with RCCs at storage days 2 and 43.
The 5%-HCT model restored the 2,3-DPG level and maintained the ATP level. Furthermore, glucose consumption and corresponding lactate production were increased in the 5%- vs the 30%-HCT condition. Lower hemolysis was observed with 5%-HCT, but only at day 2. However, morphological analysis by digital holographic microscopy (DHM) revealed a decreased fraction of discocytes at 5% rather than at 30% of HCT at storage day 2 but at day 43, the trend was inverted. Concordantly, RBCs incubated at 5% of HCT were more deformable than at 30% at day 43 (p<0.0001).
Higher metabolic activity of RBCs in the 5%-HCT condition was promoted by a higher glucose availability and limited cell-waste accumulation. The conditions of the new proposed model thus enabled rejuvenation of RBCs and maintained them in a physiological-close state in contrast to the 30%-HCT model. It may be used as a first approach to evaluate e.g., the impact of donor and recipient characteristics on RBC properties.
红细胞（RBC）在红细胞浓缩物（RCC）中的质量受到处理、储存和供体特征的影响，并可能对接受输血的患者产生临床影响。为了评估 RBC 的特性及其在输血环境中的潜在影响，已经开发了一种简单的体外输血模型。
通过将来自两个男性来源的 RCC 的洗涤 RBC 池与 15 个男性来源的新鲜冷冻血浆（FFP）单位的池在 37°C、5%CO 下混合，以 30%（“对照”设置）或 5%（替代模型）的血细胞比容（HCT）进行输血模拟。混合物孵育 48 小时。在几个孵育时间点测量不同的代谢物、溶血和微泡（MV），并在孵育后测量 RBC 形态变化和变形性。对于每个模型，在储存第 2 天和第 43 天使用 RCC 对生物学重复进行了 3 次研究。
5%-HCT 模型恢复了 2,3-DPG 水平并维持了 ATP 水平。此外，与 30%-HCT 条件相比，5%-HCT 条件下葡萄糖消耗和相应的乳酸产量增加。在 5%-HCT 条件下观察到较低的溶血，但仅在第 2 天。然而，通过数字全息显微镜（DHM）进行的形态分析显示，在第 2 天储存时，与 30%HCT 相比，盘状细胞的比例降低，但在第 43 天，趋势相反。相应地，在第 43 天，在 5%HCT 孵育的 RBC 比在 30%HCT 孵育的 RBC 更具变形性（p<0.0001）。
在 5%-HCT 条件下，RBC 的更高代谢活性是由更高的葡萄糖可用性和有限的细胞废物积累所促进的。因此，与 30%-HCT 模型相比，新提出的模型的条件使 RBC 恢复活力并使它们保持生理接近状态。它可以用作评估供体和受体特征对 RBC 特性影响的第一种方法。
