解冻的红血球的长期储存

V. Orlyk, B. Kondratskyi, Mariya Vynarchyk, Sofia Prymak, Halyna Savuliak, Vasyl Novak
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The object of the study were RBCs that were cryopreserved at -40ºС and stored for 7 days at a temperature of +2ºС - +4ºС after thawing. Deglycerolization of the thawed red blood cells, cryopreserved at -40ºС, required three time washing by using reverse cytoagglomeration. Thawed RBCs were re-suspended in lactate-sucrose-phosphate solution. After RBC thawing and storage for 7 days (186 doses) in the suspension the following indicators were studied: free hemoglobin, extracellular potassium, adenosine triphosphoric acid (ATP), 2,3-diphosphoglycerate (2,3-DPG), hematocrit, degree of hemoglobin affinity to oxygen (P50,), viscosity coefficient, osmotic stability, electrophoretic mobility of erythrocytes. as well as the total number of cells lost and recovered. \nResults. 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引用次数: 0

摘要

介绍。在现代输血实践中,无论是在和平时期还是在军事条件下,红细胞(rbc)都被广泛用作献血者血液的主要成分。冷冻保存的红细胞被认为是最安全和高质量的含红细胞环境。然而,冷冻保存后解冻红细胞的保存时间限制为24小时,这大大增加了其使用的复杂性。因此,延长解冻红细胞的保存期限对血液服务具有重要意义。研究目的:研究在-40ºС低温保存,解冻后在+2ºС - +4ºС温度下保存7天的红细胞形态状态和功能完整性。材料和方法。研究对象是在-40ºС低温保存的红细胞,解冻后在+2ºС - +4ºС的温度下保存7天。解冻后的红细胞在-40ºС低温保存,脱甘油需要用反向细胞团聚法三次洗涤。解冻后的红细胞重新悬浮在乳酸-蔗糖-磷酸溶液中。红细胞在悬浮液中解冻储存7天(186剂)后,研究以下指标:游离血红蛋白、细胞外钾、三磷酸腺苷(ATP)、2,3-二磷酸甘油酸(2,3- dpg)、红细胞压积、血红蛋白对氧亲和力(P50)、粘度系数、渗透稳定性、红细胞的电泳迁移率。以及丢失和恢复的细胞总数。结果。解冻后的红细胞在+2ºС - +4ºС温度下悬浮保存7天后,游离血红蛋白(0.62±0.02 g/l)、细胞外钾(2.7±0.3 mmol/l)、红细胞压积(0.4±0.02 l/l)等指标均在正常范围内。红细胞的渗透阻力(0.46±0.02%)、电泳迁移率(0.94±0.04µm·cm·V-1·s-1)、悬浮粘度因子(5,5±0.20 mpa·С)与正常值相比无明显变化。ATP指标(3,0±0.2µmol/gHb)和2,3- dpg指标(10,5±1,3µmol/gHb)均达到高水平。指标Р50(24.1±1.3 hPa)对应血红蛋白对氧的亲和力较低。在+2ºС - +4ºС条件下保存7天后,细胞总损失不显著,为5.6±0.4%。解冻后红细胞存活率为94,4±0,5%。结论。在-40ºС低温保存的解冻红细胞,通过反向细胞团聚脱甘油,以及使用乳酸-蔗糖-磷酸溶液对洗涤后的红细胞进行重悬,可延长解冻后的红细胞在+2ºС - +4ºС的存活条件下的储存期,最长可达7天。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
PROLONGED STORAGE OF THAWED RED BLOOD CELLS
Introduction. In modern transfusion practice, both in peacetime and in military conditions, red blood cells (RBCs) are widely used as the main component of donor blood. Cryopreserved red blood cells are considered the most safe and high-quality RBC-containing environment. However, the storage period of thawed RBCs after cryopreservation is limited to 24 hours, and significantly complicates their use. Therefore, extending the storage period of thawed RBCs is relevant for the blood service. Research objective: study the RBCs morphological state and functional completeness that were cryopreserved at -40ºС and stored for 7 days at a temperature of +2ºС - +4ºС after thawing. Materials and methods. The object of the study were RBCs that were cryopreserved at -40ºС and stored for 7 days at a temperature of +2ºС - +4ºС after thawing. Deglycerolization of the thawed red blood cells, cryopreserved at -40ºС, required three time washing by using reverse cytoagglomeration. Thawed RBCs were re-suspended in lactate-sucrose-phosphate solution. After RBC thawing and storage for 7 days (186 doses) in the suspension the following indicators were studied: free hemoglobin, extracellular potassium, adenosine triphosphoric acid (ATP), 2,3-diphosphoglycerate (2,3-DPG), hematocrit, degree of hemoglobin affinity to oxygen (P50,), viscosity coefficient, osmotic stability, electrophoretic mobility of erythrocytes. as well as the total number of cells lost and recovered. Results. After storage for 7 days of suspension of thawed RBCs at a temperature of +2ºС - +4ºС indicators of free hemoglobin (0,62±0,02 g/l), extracellular potassium (2,7±0,3 mmol/l), hematocrit (0,4±0,02 l/l) were within normal limits. Osmotic resistance (0,46±0,02%), electrophoretic mobility (0,94±0,04 µm·cm·V-1·s-1) of RBCs, suspension viscosity factor (5,5±0,20mPa·С) did not exhibit changes in comparison with normal values. High levels of ATP indicators (3,0±0,2 µmol/gHb) and 2,3-DPG (10,5±1,3 µmol/gHb) were established. Indicator Р50 (24,1±1,3 hPa) corresponded to low hemoglobine affinity for oxygen. After 7-day storage at +2ºС - +4ºС total cell loss was insignificant and amounted to 5,6±0,4%. High percentage of viable thawed RBCs 94,4±0,5% was shown. Conclusions. Deglycerolization of thawed red blood cells, cryopreserved at -40ºС, by reverse cytoagglomeration, as well as use of lactate-sucrose-phosphate solution for washed RBCs resuspending promote prolongation of thawed RBCs storage period up to 7 days at +2ºС - +4ºС in viable condition.
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