Platelet Function in Platelet Concentrates during Storage: Comparison of Two Blood Cell Separators

Latza, Reinhard; Koscielny, Jürgen; Radtke, H.; Baumann-Baretti, B.; Bläsi, Udo; Kiesewetter, H.; Jung, F.

doi:10.1159/000025250

Cited by 7 publications

(8 citation statements)

References 21 publications

(26 reference statements)

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“…In conclusion, the study showed that an increasing number of platelets became activated during 24 h of storage and could no longer be induced to adhere or to aggregate appropriately [30]. At the same time, platelets dissolved and cell debris as well as microparticles occurred.…”

Section: Resultsmentioning

confidence: 81%

Changes in platelet morphology and function during 24 hours of storage

Braune

Walter

Schulze

et al. 2014

Clinical Hemorheology and Microcirculation

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For in vitro studies assessing the interaction of platelets with implant materials, common and standardized protocols for the preparation of platelet rich plasma (PRP) are lacking, which may lead to non-matching results due to the diversity of applied protocols. Particularly, the aging of platelets during prolonged preparation and storage times is discussed to lead to an underestimation of the material thrombogenicity. Here, we study the influence of whole blood-and PRP-storage times on changes in platelet morphology and function.Blood from apparently healthy subjects was collected according to a standardized protocol and examined immediately after blood collection, four hours and twenty four hours later. The capability of platelets to adhere and form stable aggregates (PFA100, closure time) was examined in sodium citrate anticoagulated whole blood (WB) using the agonists equine type I collagen and epinephrine bitartrate (collagen/epinephrine) as well as equine type I collagen and adenosine-5 -diphosphate (collagen/ADP). Circulating platelets were quantified at each time point. Morphology of platelets and platelet aggregates were visualized microscopically and measured using an electric field multi-channel counting system (CASY). The percentage of activated platelets was assessed by means of P-selectin (CD62P) expression of circulating platelets. Furthermore, platelet factor 4 (PF4) release was measured in platelet poor plasma (PPP) at each time point.Whole blood PFA100 closure times increased after stimulation with collagen/ADP and collagen/epinephrine. Twenty four hours after blood collection, both parameters were prolonged pathologically above the upper limit of the reference range. Numbers of circulating platelets, measured in PRP, decreased after four hours, but no longer after twenty four hours. Mean platelet volumes (MPV) and platelet large cell ratios (P-LCR, 12 fL -40 fL) decreased over time. Immediately after blood collection, no debris or platelet aggregates could be visualized microscopically. After four hours, first debris and very small aggregates occurred. After 24 hours, platelet aggregates and also debris progressively increased. In accordance to this, the CASY system revealed an increase of platelet aggregates (up to 90 m diameter) with increasing storage time. The percentage of CD62P positive platelets and PF4 increased significantly with storage time in resting PRP. When soluble ADP was added to stored PRP samples, the number of activatable platelets decreased significantly over storage time.The present study reveals the importance of a consequent standardization in the preparation of WB and PRP. Platelet morphology and function, particularly platelet reactivity to adherent or soluble agonists in their surrounding milieu, changed rapidly outside the vascular system. This knowledge is of crucial interest, particularly in the field of biomaterial development for cardiovascular applications, and may help to define common standards in the in vitro hemocompatibility testing of biomaterials.

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Section: Resultsmentioning

confidence: 81%

Changes in platelet morphology and function during 24 hours of storage

Braune

Walter

Schulze

et al. 2014

Clinical Hemorheology and Microcirculation

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“…The contents of the syringe are well mixed during the blood withdrawal. [2][3][4][5][6] Within 30 minutes after blood sampling and careful homogenization, the mixture is assayed in an automatic hematology analyzer (Sysmex M-2000, Digitana AG, Hamburg, Germany). The stability of test samples is at least 2 hours.…”

Section: Methodsmentioning

confidence: 99%

“…The stability of test samples is at least 2 hours. [4][5][6] The number of platelets in the EDTA-formalin sample is reduced by the number of platelets forming as platelet aggregates. Using the ratio of the red blood cell count of both samples (EDTA / EDTA-formalin) the measurement error arising from the blood sampling can be corrected.…”

Section: Methodsmentioning

confidence: 99%

“…This is represented in a higher ratio and indicates platelet aggregation in vivo. The PR by Grotemeyer [2][3][4][5][6] Considering accurate preanalytic conditions 7 the coefficient of variation was in serial determinations 2.5%, and 1.3% in day-to-day determinations. [4][5][6] Actually, a set of sample monovettes for the PR costs 3.50 s (Rheomed, Hiddenhausen, Germany).…”

Section: Methodsmentioning

confidence: 99%

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Use of the Platelet Reactivity Index by Grotemeyer, Platelet Function Analyzer, and Retention Test Homburg To Monitor Therapy with Antiplatelet Drugs

Koscielny

Aslan

Meyer

et al. 2005

Semin Thromb Hemost

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In 1974, Wu and Hoak described a method for determining circulating platelet aggregates. This method was modified by Grotemeyer in 1983. The platelet reactivity index (PR) is based on the ratio of platelet aggregates in blood samples obtained in different buffer solutions. Platelet aggregates are resolved when blood is sampled in EDTA-buffer, but remain fixed when EDTA-formalin-buffer is used. Generally, the PR is preferred, because in vitro manipulations of platelets are not necessary, and the results are calculated. PR values above 1.05 are suspicious for elevated platelet aggregation. PR values above 1.2 indicate pathological changes in platelet aggregation. The PR is inexpensive (4.0 euro dollars) and rapid to perform. PR values were used successfully to identify nonresponders to secondary prophylaxis with acetylsalicylic acid (ASA), that is, patients suffering from stroke (33%) and patients after cardiac ischemia (18%). Furthermore, elevated PR values correlated significantly with the incidence of arterial thromboembolic complications. The PR correlated well in our prospective study with values received from the retention test Homburg (RT-H) and the platelet function analyzer (PFA-100). The data indicate that the values of the PR seem to be highly predictive for the evaluation of the ASA therapy. However, the PR is not feasible for the determination of the ASA overdosage.

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“…The selection of blood from animal species (e.g., pig and sheep) instead of human blood has to be avoided, particularly in view of the species‐dependent platelet function . In blood products, such as frozen plasma or platelet concentrates, proteins (e.g., factors V and VIII, or cytokines) and cells (e.g., platelet adhesion and aggregation) undergo functional alterations due to the processing (e.g., cooling and freezing), storage (e.g., storage lesion of cells), and stabilization . Thus, the use of blood products should be avoided since it very likely lead to an underestimation of the thrombogenicity of the tested materials.…”

Section: Prerequisites For a Reproducible In Vitro Testingmentioning

confidence: 99%

In Vitro Thrombogenicity Testing of Biomaterials

Braune

Latour

Reinthaler

et al. 2019

Adv Healthcare Materials

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reliable, and reproducible. [20][21][22] This is, not least, important for realizing interstudy comparisons of the thrombogenicity of different materials. According, e.g., to the regulation (EU) 2017/745 of the European parliament and of the council on medical devices, in vitro and in vivo tests are a mandatory parts within the preclinical product verification and validation (see Annex II: Technical documentation, 6.1 Preclinical and clinical data). [23] Beyond the results of these tests, also detailed information concerning the test design, respective protocols and data analysis are obligatory to meet the requirements of the regulation, particularly those formulated in the ISO standard 10993 Part 4. [24,25] Since the regulatory agencies provide recommendations rather than protocols or standard operation procedures, accepted standardizations regarding the preanalytical handling of the blood, test conditions, test parameters, reference materials, static or dynamic conditions, flow conditions, and finally, which type of cells and donors should be included is lacking. Also, standard operating procedures how the testing should be performed are lacking. [9] This has led to a situation that laboratories perform in vitro assays under substantially different test conditions, including the preanalytical characterization of blood samples, the application of blood from different species, varying anticoagulation and stabilization or storage times as well as test setups. Taking these variations into account, interlaboratory or interstudy comparisons are impossible. [8,[26][27][28] Here, we review described test methods and procedures for the assessment of the thrombogenicity of materials, prerequisites for a reproducible in vitro testing, applied test system and test parameters for the characterization of the interaction of blood components/cells and the implant. We further review and outline recent approaches that may support realizing reliable, reproducible, and laboratory independent tests.

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Platelet Function in Platelet Concentrates during Storage: Comparison of Two Blood Cell Separators

Cited by 7 publications

References 21 publications

Changes in platelet morphology and function during 24 hours of storage

Changes in platelet morphology and function during 24 hours of storage

Use of the Platelet Reactivity Index by Grotemeyer, Platelet Function Analyzer, and Retention Test Homburg To Monitor Therapy with Antiplatelet Drugs

In Vitro Thrombogenicity Testing of Biomaterials

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