: AAbstractObjective:To compare and analyze the metabolic and functional changes in platelets stored at 4 ℃ and ones stored at 22 ℃ with agitation so as to provide an experimental basis for the cryopreservation technology of platelets. METHODS: Samples were collected from platelets stored at 4 ℃ in 2, 4, 6, 11, 15 and 21 days, and from ones stored at 22 ℃ with agitation during the same days, the metabolism indicators and thromboelastogram (TEM) were analysed. RESULTS: In metabolism, there were no significant changes of pH, GLU，PCO2, PCO2 and MPV levels of platelets stored at 4 ℃ for <6 days (P＞0.05), However, the Plt count decreased, the PDW and LDH level incrseased (P＜0.05). At the same time, only MPV had no changes of platelets stored at 22 ℃ during above-mentioned same days (P＞0.05), while the pH, PCO2, GLU, Plt all decreased, and PO2, LDH, PDW incrseased (P＜0.05). There were significant changes about the pH value, PO2, Plt, MPV, LDH, GLU levels between the two kinds of stored platelets during the same storing period (P＜0.05). The pH value and MPV of platelets stored at 4 ℃ were obviously lower than ones stored at 22 ℃, while GLU, PO2, LDH and Plt levels showed reverse changes (P＜0.05). Meanwhile, the PCO2 of platelets stored at 4 ℃ not could be detected and the Plt count reduced rapidly from d15. In function, the MA level of platelets stored at 4 ℃ was slower than that of platelets stored at 22 ℃, that is, the MA level of platelets stored at 4 ℃ were higher than that of platelets stored at 22 ℃ during the same storeing period (P＜0.05). CONCLUSION Platelets stored at 4 ℃ have much slower metabolism than ones stored at 22 ℃, and the aggregation is stronger of platelets stored at 4 ℃ than that of ones at 22 ℃ during the same conservation period.
Blood Platelets ; Blood Preservation ; Cryopreservation ; Temperature

The objective of the present study was designed to evaluate lyophilized red blood cells of the ultrastructure. Blood was drawn from healthy adult. In group 1, sample was fresh blood; in group 2, sample was added 35% glycerine, stored at -80 degrees C for 24 hours; in group 3, red blood cells stored at 4 degrees C for 5 hours, then were lyophilized for 16 hour. The sample was resuspended for measurements of count and electron microscopy study. The result showed that lyophilized red blood cells possessed relative integrated structure, red blood cell recovery was 53%. The mean diameter, optical density and integral optical density of red blood cell were 4.7 +/- 0.4, 0.14 +/- 0.03 and 1.58 +/- 0.46 in group 1; 4.6 +/- 0.7, 0.14 +/- 0.02 and 2.35 +/- 0.64 in group 2; 4.4 +/- 0.4, 0.17 +/- 0.05 and 2.35 +/- 0.46 in group 3, respectively. There was no significant difference in lyophilized and frozen group, but there was significant difference in lyophilized group and normal group. In conclusion, human red blood cells could be successfully lyophilized and possess relative integrated structure. The mean diameter, optical density and integral optical density of lyophilized red blood cells were similar to that of cryopreservation red cells.
Blood Preservation ; Erythrocytes ; ultrastructure ; Freeze Drying ; Humans

The temperature of platelet intracellular ice crystal formation (IIF) is one of the most important physical parameters to instruct platelet cryopreservation. In this study, the range of temperatures for platelet IIF was measured by means of biological and physical methods. All platelet samples were graded cooling, and two samples of per 5 degrees C decrease were thawed by 2 different ways: 37 degrees C directly (T 37 degrees C) and 37 degrees C after keeping in liquid nitrogen (LN) for 2 hours. The phosphatidylserine (PS) positive rate, plasma lactate dehydrogenase (LDH) concentration and platelet aggregate rate were measured in all samples. The heat release graphs of platelets cryopreserved with or without 5% DMSO were also measured by differential scanning calorimeter (DSC). The results showed that the PS positive rates and aggregate rates in platelets and plasma LDH concentrations gradually increased in T 37 degrees C group and decreased in LN group until the arrival of -35 degrees C, and then there were no further changes of the 3 parameters. A small second heat release peak was detected at about -35 degrees C in the platelet samples cryopreserved without DMSO. It is concluded that the temperature of intracellular ice crystal formation in platelet is from -30 to -40 degrees C (-35 degrees ).
Blood Platelets ; physiology ; Blood Preservation ; Cryopreservation ; Crystallization ; Humans ; Temperature

Platelets are important component in the blood which play vital role in physiological hemostasis. In clinic, platelets are usually transfused to patients suffered from idiopathic thrombocytopenic purpura,thrombocytopenia after chemotherapy and radiotherapy,or hemorrhagic thrombocytopenia caused by other reasons. Storage time for platelet concentrates is limited to five days at room temperature, leading to an increasing number of scrap rate, therefore the prolonging the storage time is important for clinical application. Storage at -80°C is an ideal long-term cryopreservation method for platelet concentrates. In this article, platelets cryopreservation,the mechanism of frozen injury and the protective mechanisms of platelets were reviewed.
Blood Platelets ; Blood Preservation ; Cryopreservation ; Freezing ; Hemostasis ; Humans ; Thrombocytopenia

OBJECTIVETo evaluate the storage performance of the domestically made platelet storage bags (experimental group) and the United States Trima set platelet storage bags (control group).

METHODSThe manually separated platelets were divided in two equal parts, which was added to control blood bags and experimental blood bags respectively, all samples were stored at a 22 °C ± 2 °C. The platelet count, mean volume, aggregation activity (ADP, THR), pH, glucose, lactate concentration, lactate dehydrogenase concentration, hypotonic shock reaction, CD62P and phosphatidic acid serine content were detected at day 0, 3, 5 and 7 of storage.

RESULTSThere was no significant difference of platelet quality at day 5 after storage between the experimental group and the control group (T-test, P > 0.05).

CONCLUSIONTwo kinds of platelet storage bags have the similar storage performance.

Chemokine is a class of soluble active peptides that attract white blood cells to the inflammatory site. CD40 ligand (CD40L) involves in synthesis of proinflammatory mediators. Accumulation of chemokine and CD40L can induce non-hemolytic reaction after transfusion, transfusion-related acute lung injury (TRALI) and autoimmune disease during blood component storage. Pre-storage leucocyte deletion can prevent the release of leucocyte-derived chemokines, but not prevent the accumulation of platelet-derived chemokines. γ-irradiation or ultraviolet β irradiation is effective in preventing the increase of chemokines in the storage of platelet, thus prevents non-hemolytic febrile reaction after platelet transfusion. In this review, the recent advance in research of accumulation of chemokines and CD40L during blood component storage, and its effect on blood transfusion, as well as preventive measures are summarized.
Blood Platelets ; metabolism ; Blood Preservation ; Blood Transfusion ; CD40 Ligand ; blood ; Chemokines ; blood ; Humans

Now the clinical preservation methods of human red blood cells mainly include hypothermic storage (4 degrees C) and cryopreservation (-80 degrees C or -196 degrees C). The preservation time of hypothermic storage of red blood cells is relatively short and it is easy to be contaminated by microbes. Cryopreservation greatly prolongs the storage time, but it needs heavy storage equipments. Because the protective solutions in cryopreservation contain glycerol, red blood cells need complicated washing in order to remove glycerol. These shortage methods limit their application to some special conditions, such as war or natural disasters. Compared with conventional preservation methods of red blood cells, lyophilization has many advantages such as less weight, convenient transportation, room temperature preservation, prone to be rehydrated. In this review, the progress and challenge in the development of lyophilization of red blood cells, especially application of trehalose and its mechanism in the lyophilization of red blood cells were systematically discussed. This review can provide some theoretic guidance for developing a safe, simple and efficient preservation approach of red blood cells by lyophilization.
Blood Preservation ; Cryopreservation ; Erythrocytes ; Freeze Drying ; Humans ; Trehalose ; pharmacology

To study effect of pre-freezing temperature and lyophilizer shelf temperature on recovery of human red blood cells after lyophilization and determine solidifying temperature of this lyophilization system, the protective solution composed of 7% DMSO, 40% polyvinylpyrrolidone (PVP) and isotonic buffer were adopted to lyophilize red blood cells at different pre-freezing temperatures or shelf temperatures. At first, fresh whole blood was centrifugated, washed and equilibrized to prepare concentrated red blood cells. Then concentrated red blood cells were mixed with the protective solution at 1:3 and pre-freezed at different temperature (-20, -35, -45, -80 or -196 degrees C) before lyophilization in lyophilizer. To study effect of shelf temperature on lyophilization of red blood cells, red blood cells were lyophilized at different shelf temperature after pre-freeze at -80 degrees C. After lyophilization, the samples were quickly rehydrated by 37 degrees C rehydration solution. The results showed the recovery rate of red blood cells and hemoglobin after pre-freeze at different temperature and lyophilization were > 85% and > 75%, there was not significant difference among these groups, but the concentration of free hemoglobin in -196 degrees C group was significantly higher than that in other groups (P < 0.01). With decreasing of shelf temperature, the lyophilizing time was also prolonged. When shelf temperature was > or = -25 degrees C, samples were not fully lyophilized; when shelf temperature was < or = -30 degrees C, the recovery rate of red blood cells and hemoglobin after lyophilization and rehydration were above 90%; after washed to isotonic state, the recovery rate of hemoglobin of the four groups was similar to each other. In conclusion, only when pre-freezing temperature is between -20 and -80 degrees C and the lyophilizer shelf temperature is < or = -30 degrees C, the effect of lyophilization is better, but the effect of excessively low pre-freezing temperature may even be worse.
Blood Preservation ; Erythrocytes ; cytology ; Freeze Drying ; Hemoglobins ; analysis ; Humans ; Temperature

OBJECTIVE: To explore the metabolic characteristics and metabolic markers of WBC-depleted RBCs in MAP preservation solution and to analyzed the metabolic profile of RBC in MAP preservation solution by using metabolomics. METHODS: The changes of metabolitcs in 10 U WBC-depleted RBC suspension at 3-different storage period (D 0, D 14 and D 35) were detected by using the UPLC-MS/MS, the charaeteristic ions and metabolic markers of RBC stored in preservation sblution for 0 d, 14 d and 35 days were analyzed by using the principal component analysis(PCA). RESULTS: The number of characteristic ions in RBC and supernatant extracts detected during the initial, middle and final storage could be clearly distinguiseed. The 5 metabolism-related substamces such as lact-c acid, nicotinamide, glucose, 5-htdroxyproline and malic acid showed statistically significant difference in 3 storage period. CONCLUSION The UPLC-MS/MS method combined with statistical analysis of multivariate data can be used to study the metabolic characteristics of RBC, the different metabolites of RBC in different storage stages can be used as the potential markers for evaluation of guality of RBC in storag period. The results of this study provide a basis for studing the RBC guality changes in storage period.
Blood Preservation ; Chromatography, Liquid ; Erythrocytes ; Metabolome ; Tandem Mass Spectrometry

OBJECTIVETo investigate the differences of metabolic pathways of leucocyte-deplated RBCs prepared by using lipid whole blood and nomal blood during routine storage so as to provide some reference for clinical blood use.

METHODSTwenty U whole blood from 20 donors, including 10 U lipid blood and 10 U normal whole blood, were selected for preparing leukodepleted red blood cells, red blood cells were taken from storage bags on day 0, 14 and 35, respectively. Metabolites in the red blood cells were analyzed, red blood cell metabolic extracts were detected by UPLC-MS/MS. The metabolite data of RBC from 2 groups were analyzed by SIMCA-P 13.0 software using OPLS-DA and by SPSS 19.0 using Mann-Whitney U test. Difference of metabolic pathways was described according to different metabolites.

RESULTSThe glucose, adenine, pyruvic acid, GSH, GSSG and niacinamide levels on day 0 in lipid RBCs were higher than those in the control group(P<0.05). The glucose, pyruvic acid and GSH levels on day 14 in lipid RBCs were lower than those in the control group (P<0.05), and the levels of adenine, GSSG and niacinamide were higher than that in the control group (P<0.05). The glucose level on day 0 was lower than that in the control group (P<0.05), and the levels of adenine and niacinamide were higher than those in the control group (P<0.05). but the pyruvic acid, GSH and GSSG levels were not significantly different between 2 groups (P>0.05).

CONCLUSIONCompared with the normal red blood cells, the energy metabolism pathway decreases in lipid red blood cells within the storage period and pentose phosphate pathway increases.