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Blood Mar 2013Assessing messenger RNA (mRNA) and microRNA levels in peripheral blood cells may complement conventional parameters in clinical practice. Working with small, precious...
Assessing messenger RNA (mRNA) and microRNA levels in peripheral blood cells may complement conventional parameters in clinical practice. Working with small, precious samples requires optimal RNA yields and minimal RNA degradation. Several procedures for RNA extraction and complementary DNA (cDNA) synthesis were compared for their efficiency. The effect on RNA quality of freeze-thawing peripheral blood cells and storage in preserving reagents was investigated. In terms of RNA yield and convenience, quality quantitative polymerase chain reaction signals per nanogram of total RNA and using NucleoSpin and mirVana columns is preferable. The SuperScript III protocol results in the highest cDNA yields. During conventional procedures of storing peripheral blood cells at -180°C and thawing them thereafter, RNA integrity is maintained. TRIzol preserves RNA in cells stored at -20°C. Detection of mRNA levels significantly decreases in degraded RNA samples, whereas microRNA molecules remain relatively stable. When standardized to reference targets, mRNA transcripts and microRNAs can be reliably quantified in moderately degraded (quality index 4-7) and severely degraded (quality index <4) RNA samples, respectively. We describe a strategy for obtaining high-quality and quantity RNA from fresh and stored cells from blood. The results serve as a guideline for sensitive mRNA and microRNA expression assessment in clinical material.
Topics: Algorithms; Blood Cells; Blood Chemical Analysis; Blood Preservation; Blood Specimen Collection; Calibration; Genetic Techniques; Guanidines; Humans; MicroRNAs; Phenols; Polymerase Chain Reaction; Preservation, Biological; RNA Stability; RNA, Messenger; Reference Standards
PubMed: 23327925
DOI: 10.1182/blood-2012-06-438887 -
Nature Communications Oct 2022Deoxyribonucleic acid (DNA) is the blueprint of life, and cost-effective methods for its long-term storage could have many potential benefits to society. Here we present...
Deoxyribonucleic acid (DNA) is the blueprint of life, and cost-effective methods for its long-term storage could have many potential benefits to society. Here we present the method of in situ cryosilicification of whole blood cells, which allows long-term preservation of DNA. Importantly, our straightforward approach is inexpensive, reliable, and yields cryosilicified samples that fulfill the essential criteria for safe, long-term DNA preservation, namely robustness against external stressors, such as radical oxygen species or ultraviolet radiation, and long-term stability in humid conditions at elevated temperatures. Our approach could enable the room temperature storage of genomic information in book-size format for more than one thousand years (thermally equivalent), costing only 0.5 $/person. Additionally, our demonstration of 3D-printed DNA banking artefacts, could potentially allow 'artificial fossilization'.
Topics: Humans; Ultraviolet Rays; DNA; Blood Preservation; Preservation, Biological; Oxygen
PubMed: 36270991
DOI: 10.1038/s41467-022-33759-y -
Journal of Immunological Methods Jan 2020A potential hurdle when applying mass cytometry to the field study setting is the streamlining of sample collection while at the same time protecting the integrity of... (Comparative Study)
Comparative Study
A potential hurdle when applying mass cytometry to the field study setting is the streamlining of sample collection while at the same time protecting the integrity of important cell epitopes. Whole blood preservation kits applying fixation and/or permeabilization agents are increasingly used in clinical trials to preserve leukocytes needed for downstream analysis. We here present a structured overview of leukocyte surface marker detectability in samples processed with four commercially available whole blood preservation kits; 1) Proteomic Stabilizer, 2) Stable-Lyse V2 and Stable-Store V2, 3) Cytodelics and 4) Lyse/Fix Buffer, as well as in samples treated with buffers included in Mass-tag Cellular Barcoding kits. Isolated leukocytes were stained with a 28-marker panel (including 7 chemokine receptors) of metal-conjugated antibodies and analysed on a mass cytometer. Exploration of the data by manual gating and viSNE analysis showed that although many markers were similarly detected across all sample conditions, most of the chemokine receptors in our panel, particularly CXCR3, CCR4, CCR6 and CXCR5, were incorrectly detected in the preserved samples and thus incompatible with the fixation and permeabilization agents found in whole blood preservation kits and in buffers used prior to barcoding.
Topics: Adult; Antigens, CD; Blood Preservation; Fixatives; Flow Cytometry; Humans; Leukocytes; Receptors, Chemokine
PubMed: 31629741
DOI: 10.1016/j.jim.2019.112673 -
Journal of Veterinary Emergency and... Nov 2022Despite a lack of strong evidence of benefit, leukoreduction is employed to decrease the risk of leukocyte-induced transfusion reactions. However, the impact of...
BACKGROUND
Despite a lack of strong evidence of benefit, leukoreduction is employed to decrease the risk of leukocyte-induced transfusion reactions. However, the impact of leukoreduction on blood bank costs and inventory management is not well understood. The purpose of this study was to determine whether leukoreduction of whole blood increases total processing time and weight loss from packed red blood cells (PRBCs) and plasma relative to bags created from nonleukoreduced whole blood.
KEY FINDINGS
A total of 68 canine whole blood collections were divided equally into leukoreduced and nonleukoreduced groups (N = 34 in each). There was no significant difference between groups in mean PRBC or plasma unit weights or processing times. Leukoreduced PRBC bags lost a significantly greater proportion of weight during processing than did nonleukoreduced PRBC bags (P < 0.01), which is attributed to red and white blood cells lost in the filtration process.
SIGNIFICANCE
Leukoreduction did not lead to a significant increase in processing times or smaller PRBCs or plasma bags compared to nonleukoreduced bags. The blood remaining in the leukoreduction filter following filtration is primarily composed of red blood cells, with minimal plasma retained.
Topics: Dogs; Animals; Erythrocyte Transfusion; Erythrocytes; Leukocytes; Blood Banks; Leukocyte Count; Blood Preservation
PubMed: 35712893
DOI: 10.1111/vec.13225 -
International Journal of Molecular... Jan 2022Cold-stored platelets are making a comeback. They were abandoned in the late 1960s in favor of room-temperature stored platelets due to the need for longer... (Review)
Review
Cold-stored platelets are making a comeback. They were abandoned in the late 1960s in favor of room-temperature stored platelets due to the need for longer post-transfusion platelet recoverability and survivability in patients with chronic thrombocytopenia. However, the current needs for platelet transfusions are rapidly changing. Today, more platelets are given to patients who are actively bleeding, such as ones receiving cardiac surgeries. It has been established that cold-stored platelets are more hemostatically effective, have reduced bacterial growth, and have longer potential shelf lives. These compelling characteristics led to the recent interest in bringing back cold-stored platelets to the blood systems. However, before reinstating cold-stored platelets in the clinics again, a thorough investigation of in vitro storage characteristics and in vivo transfusion effects is required. This review aims to provide an update on the recent research efforts into the storage characteristics and functions of cold-stored platelets using modern investigative tools. We will also discuss efforts made to improve cold-stored platelets to be a better and safer product. Finally, we will finish off with discussing the relevance of in vitro data to in vivo transfusion results and provide insights and directions for future investigations of cold-stored platelets.
Topics: Animals; Blood Platelets; Blood Preservation; Cryopreservation; Humans; Platelet Transfusion
PubMed: 35163024
DOI: 10.3390/ijms23031100 -
Cell Proliferation Apr 2011To review information on cord blood banking and transplantation with respect to the author's studies, and in context of this field of investigation. (Review)
Review
OBJECTIVES
To review information on cord blood banking and transplantation with respect to the author's studies, and in context of this field of investigation.
RESULTS
Cord blood transplantation has been successfully used to treat a number of malignant and non-malignant disorders. However, this technique is still associated with limited numbers of cells for transplantation, and with delayed engraftment of neutrophils and platelets. The field of cord blood transplantation will benefit from enhanced and mechanistically based information on haematopoietic stem cell function and potential means to enhance its effectiveness are reviewed. This includes notions concerning possibility of retrieving more cells from the placenta and cord blood, to expand haematopoietic stem cells ex vivo and to increase efficiency of homing and engraftment of these cells. Also discussed are cryopreservation and long-term storage of cord blood haematopoietic and progenitor cells, and new laboratory findings and animal studies for non-haematopoietic uses of cord blood.
Topics: Animals; Blood Banking; Blood Preservation; Cord Blood Stem Cell Transplantation; Fetal Blood; Hematopoietic Stem Cell Transplantation; Humans
PubMed: 21481045
DOI: 10.1111/j.1365-2184.2010.00728.x -
Journal of Veterinary Internal Medicine Mar 2021Leukoreduction is a routine procedure in human transfusion medicine but is uncommon in veterinary.
BACKGROUND
Leukoreduction is a routine procedure in human transfusion medicine but is uncommon in veterinary.
OBJECTIVES
To evaluate the effect of leukoreduction on the quality of canine whole blood (WB) and blood products during storage.
ANIMALS
Ten canine blood donors.
METHODS
This is a case series study. An amount of 450 mL of blood was collected from each dog. Five WB and 5 packed red blood cells (pRBC) bags were divided into 2 units each: leukoreduced (LR) and non-leukoreduced (nLR). RBC count, erythrocytes' mean osmotic fragility (MOF), 2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), percentage of hemolysis, potassium (K), lactate, glucose, and cytokines were measured weekly from day of donation (T0) to day 35 (T35); pH, coagulation times, and clotting factors were evaluated at T0 and T35 from WB and in fresh frozen plasma after 1 year of storage.
RESULTS
Leukoreduction showed positive effects on lactate (T35: LR WB 14.42 mmol/L SD 2.71, nLR WB 22.42 mmol/L SD 1.86, LR pRBC 20.88 mmol/L SD 2.65, nLR pRBC 36.81 mmol/L SD 2.34; P < .0001), pH (T35: LR WB 6.88 SD 0.16, nLR WB 6.69 SD 0.20, P = .02; LR pRBC 6.57 SD 0.23, nLR pRBC 6.22 SD 0.11; P < .001), and K (LR pRBC 4.08 mmol/L SD 0.88, nLR pRBC 5.48 mmol/L SD 0.90; P < .001). Increasing values of IL8 were observed in nLR units during storage (T0: 4167 ± 11 888 pg/mL; T35: 6367 ± 11 612 pg/mL).
CONCLUSION AND CLINICAL IMPORTANCE
LR blood units are recommended to critically ill dogs with marked inflammatory conditions.
Topics: Adenosine Triphosphate; Animals; Blood Preservation; Cytokines; Dog Diseases; Dogs; Erythrocytes; Hemolysis
PubMed: 33591603
DOI: 10.1111/jvim.16039 -
Blood Transfusion = Trasfusione Del... May 2017Haemolysis occurs in many haematologic and non-haematologic diseases. Transfusion of packed red blood cells (pRBCs) can result in intravascular haemolysis, in which the... (Review)
Review
Haemolysis occurs in many haematologic and non-haematologic diseases. Transfusion of packed red blood cells (pRBCs) can result in intravascular haemolysis, in which the RBCs are destroyed within the circulation, and extravascular haemolysis, in which RBCs are phagocytosed in the monocyte-macrophage system. This happens especially after RBCs have been stored under refrigerated conditions for long periods. The clinical implications and the relative contribution of intra- vs extra-vascular haemolysis are still a subject of debate. They have been associated with adverse effects in animal models, but it remains to be determined whether these may be involved in mediating adverse effects in humans.
Topics: Animals; Blood Preservation; Erythrocyte Transfusion; Erythrocytes; Hemolysis; Humans
PubMed: 28518048
DOI: 10.2450/2017.0311-16 -
Blood Transfusion = Trasfusione Del... Mar 2017What is following the impressive progress that has been made? During the last couple of years several tremors have shaken the field of Transfusion Medicine. The... (Review)
Review
What is following the impressive progress that has been made? During the last couple of years several tremors have shaken the field of Transfusion Medicine. The epicentres of those tremors were located on novel insights into the RBC storage lesion, on emerging connections between storage lesion and post-transfusion performance and effects, and on acknowledging that storage time is only one (rather than the most prominent) of the parameters which contribute to the progression of storage lesion in any given unit of blood. The optimisation of bio-preservation conditions emerged at the same time with all-new scientific knowledge gained by advances in research tools, implementation of technological innovations, and application of elegant in vitro and in vivo models of transfusion. Simultaneously, one after another, all the reported randomised clinical trials concluded, with spectacular consensus, that there is no significant difference in the rate of adverse clinical events (including death) among patients who underwent transfusion with fresh (and presumably good) or standard of care (and presumably bad) blood. The comparative analysis and comprehension of the aforementioned data would set the context for the next generation of research in blood transfusion science, since the need for safer and more efficient transfusions remains.
Topics: Blood Preservation; Erythrocyte Transfusion; Humans; Randomized Controlled Trials as Topic; Time Factors
PubMed: 28263169
DOI: 10.2450/2017.0313-16 -
Seminars in Perinatology Aug 2012Red blood cell (RBC) transfusion is indicated to improve oxygen delivery to tissue, and for no other purpose. We have come to appreciate that donor RBCs are... (Review)
Review
Red blood cell (RBC) transfusion is indicated to improve oxygen delivery to tissue, and for no other purpose. We have come to appreciate that donor RBCs are fundamentally altered during processing and storage in a manner that both impairs oxygen transport efficacy and introduces additional risk by perturbing both immune and coagulation systems. The protean biophysical and physiological changes in RBC function arising from storage are termed the "storage lesion;" many have been understood for some time; for example, we know that the oxygen affinity of stored blood rises during the storage period and that intracellular allosteric regulators, notably 2,3-bisphosphoglyceric acid and ATP, are depleted during storage. Our appreciation of other storage lesion features has emerged with improved understanding of coagulation, immune, and vascular signaling systems. Here, we review key features of the "storage lesion." Additionally, we call particular attention to the newly appreciated role of RBCs in regulating linkage between regional blood flow and regional O(2) consumption by regulating the bioavailability of key vasoactive mediators in plasma, and discuss how processing and storage disturb this key signaling function and impair transfusion efficacy.
Topics: Blood Preservation; Erythrocyte Transfusion; Erythrocytes; Humans; Nitric Oxide; Oxidative Stress; Oxygen Consumption; Regional Blood Flow; Signal Transduction; Time Factors
PubMed: 22818545
DOI: 10.1053/j.semperi.2012.04.005