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Platelet Storage and Functional Integrity J Lab Med 2020; 44(5): 285–293 Review Gianmatteo Vit, Harald Klüter and Patrick Wuchter* Platelet storage and functional integrity https://doi.org/10.1515/labmed-2020-0067 Isolation of human platelets Received June 16, 2020; accepted August 23, 2020; published online September 11, 2020 Donor selection Abstract: Platelet transfusion is a topic of common in- Platelet donor recruitment underlies the German Trans- terest for many specialists involved in patient care, from fusion Act (Transfusionsgesetz) and follows the national laboratory staff to clinical physicians. Various aspects guidelines for the preparation of blood and blood compo- make this type of transfusion different from those of other nents [1]. Donors must be in a health condition that allows a blood components. In this review, the challenges in donation without risk for themselves and the recipients. platelet transfusion practice that are relevant for labo- Moreover, they must be informed in detail about the risks ratory colleagues will be discussed, highlighting how the and side effects of the donation. In addition to the general biochemical and structural characteristics of these blood prerequisites for blood donation that we will not discuss in elements directly affect their function and consequently the present review, specific requirements have to be ful- the clinical outcome. More than 1,300 platelet concen- filled for apheresis platelet donation, i.e., a platelet count trates are transfused in Germany every day, and several of ≥150,000/µL. This value is of key importance since the types are offered by their respective manufacturers. We blood platelet concentration after apheresis should be describe the technological advances in platelet concen- >100,000/µL. Drugs that impair platelet function (e.g., trate production, with a focus on how the storage con- acetylsalicylic acid [ASA] or nonsteroidal anti- ditions of platelets can be improved. Laboratory quality inflammatory drugs [NSAIDs]) must not have been assessment procedures for a safe transfusion are dis- consumed in the last seven days before donation [1]. cussed in detail. For this purpose, we will refer to the Hemotherapy Directives (Richtlinie Hämotherapie)ofthe German Medical Association. Pooled and apheresis platelet concentrates Keywords: platelet concentrate; thrombocytopenia; transfusion. Platelet concentrates can be obtained either from whole blood donations with principally two different subsequent preparation procedures or directly by apheresis technolo- gies. One option is the isolation of platelets from buffy coats (BC) derived from different, usually four to six, blood donors. To remove undesired cells (e.g., white blood cells), the centrifugation of pooled BCs is followed by a filtration *Corresponding author: Prof. (apl.) Dr. med. Patrick Wuchter, step to obtain a leukodepleted concentrate [2]. At the end of Institute of Transfusion Medicine and Immunology, Medical Faculty the procedure, the platelets are stored in platelet additive Mannheim, Heidelberg University, German Red Cross Blood Service solutions (PAS), which aim to guarantee the maintenance Baden-Württemberg - Hessen, Friedrich-Ebert-Str. 107, 68167 of the functional elements in an environment containing as Mannheim, Germany. Phone: 0621 - 3706 9581, Fax: 0621 - 3706 little plasma as possible. Furthermore, PAS provide a 9496, E-mail: [email protected] Gianmatteo Vit, Institute of Transfusion Medicine and Immunology, buffering capacity and allow few allergic reactions or Medical Faculty Mannheim, Heidelberg University, German Red Cross transfusion-associated complications [3]. Very promising Blood Service Baden-Württemberg - Hessen, Mannheim, Germany; efforts have also been made to exploit the possibility of The Novo Nordisk Foundation Center for Protein Research, Protein using buffy coats as a source of hematopoietic progenitor Signaling Program, Faculty of Health and Medical Sciences, University cells for platelet production [4]. of Copenhagen, Copenhagen, Denmark Harald Klüter, Institute of Transfusion Medicine and Immunology, Another approach is the sequential double centrifu- Medical Faculty Mannheim, Heidelberg University, German Red Cross gation of blood derived from whole blood donors. This Blood Service Baden-Württemberg - Hessen, Mannheim, Germany procedure is generally very seldom used in Europe and Open Access. © 2020 Gianmatteo Vit et al., published by De Gruyter. This work is licensed under the Creative Commons Attribution 4.0 International License. 286 Vit et al.: Platelet storage and functional integrity actually not used at all in Germany. It includes a first activation, which represents a phenomenon at the basis of centrifugation step at low speed, which separates the blood their physiological function. Platelet activation markers in two phases. In the upper phase, platelets are enriched in used in diagnosis are represented by surface molecules, the so-called platelet rich plasma (PRP). In the lower phase, such as P-selectin (CD62P, GMP-140), a receptor for white erythrocytes and white blood cells are sedimented. In a blood cells (WBCs). Under normal conditions, CD62P is second centrifugation step, the platelets harvested in the contained inside the α-granules, and the degranulation upper phase are centrifuged and subsequently recon- process is considered the initial stage of platelet activation, stituted in plasma [5]. which is followed by exposure of the receptor at the Manufacture of platelet concentrates by automated membrane. Monitoring the concentration of this receptor is apheresis procedures is widely used when platelets are of some importance, as it has been implicated in platelet collected from individual donors. With this method, clearance processes by WBCs that results in the reduction platelets are collected from a single donor through an in- in efficacy of the administered preparation [11]. Moreover, tegrated system capable of reducing the presence of leu- the expression of this receptor leads to the activation of kocytes in the final apheresis product, a process called WBCs, which in turn are able to activate macrophages, leukodepletion. The blood is withdrawn from the donor which are responsible for the release of pro-inflammatory through an apheresis needle and flows through a sterile cytokines [12]. In this way, the transfusion of activated conduit until it reaches the machine in which the platelet platelets can lead to a moderate increase in the inflam- isolation procedure takes place. Once the platelets have matory response, which may be the basis of transfusion been isolated by differential centrifugation, blood deprived reactions. However, studies have shown that P-selectin of platelets and a small amount of plasma flows back into expression at the membrane level is reversible, thus the donor. This procedure is particularly advantageous in limiting platelet clearance by WBCs [13]. Lysosomal the case of platelet refractoriness, i.e., poor platelet in- exocytosis can also be used as a read-out of platelet acti- crements after transfusion. This condition is usually due to vation by cell separators. CD63 (GP53) is a membrane antibodies present in the plasma of the donor directed protein present on lysosomes that can be monitored to against common antigens, such Human Leukocyte Antigen assess lysosomal activity [14]. Activation of the integrin I (HLA I), on the surface of leukocytes and platelets [6, 7]. complex glycoprotein IIb/IIIa (fibrinogen receptor) is a In Germany, the current guidelines demand for pooled further marker of platelet activation. This receptor medi- units as well as for apheresis platelet concentrates a min- ates both adhesion of platelets to the exposed extracellular imum number of ≥2 × 1011 platelets per unit [1]. The choice matrix and platelet cross-linking [15]. A study comparing between a pooled and an apheresis product is dictated three different types of cell separators regarding the risk of from a clinical point of view by various factors. First, in inducing platelet activation has shown that activation of many clinics, the economic factor must be considered the fibrinogen receptor is a common occurrence; in this (apheresis products are much more expensive), but the case, it was shown to be a reversible phenomenon since the clinical needs of the patient should be the guiding criterion parameter decreased to normal values after 2 h [16]. The that influences the choice. In patients who show no signs of same study also characterized the expression of glyco- immunization, there are no comparative data from pro- protein IX (CD42b), a membrane protein present on plate- spective randomized trials available [8, 9]. An undisputed lets, which constitutes the receptor of the von Willebrand indication for platelet concentrates from apheresis is the factor. A decrease in glycoprotein IX during apheresis was occurrence of HLA- or HPA-antibodies. Alloimmune unaffected after a 2 h resting period of the platelet thrombocytopenia is a pathology affecting fetuses and concentrate and a subsequent 2 h agitation [16]. newborns due to maternal antibodies directed against Although many of the structural changes following platelet antigens. These factors can impede platelet incre- platelet activation during apheresis are reversible, the ment and therefore require individually matched platelet initial lesion derived from the isolation process plays an concentrates [1, 10]. essential role in the quality of the platelet concentrate and in the efficiency of the transfusion,
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