Platelet preparation for function testing in the laboratory and clinic: Historical and practical aspects

Hechler, Béatrice; Dupuis, Arnaud; Mangin, P; Gachet, Christian

doi:10.1002/rth2.12240

Cited by 46 publications

(46 citation statements)

References 53 publications

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“…In a washed platelet suspension ( Figure 2), one does not observe the same abnormalities as in cPRP after stimulation of platelets with ADP, whatever the concentration of agonist used. This is because ADP does not induce platelet degranulation or thromboxane A2 generation in the presence of 2 mM calcium [46,51,52] and aggregation is by definition reversible. Unlike ADP-induced aggregation which remains normal, collagen (1.25 and 2.5 µg/mL)-induced aggregation generally The absence of a second wave in ADP (2.5 and 5 µM)-induced platelet aggregation; A decrease in the maximum amplitude of collagen-induced aggregation together with an increase in the lag time, especially when a low concentration of collagen is used (1.25 µg/mL) [48];…”

Section: Aggregation Testsmentioning

confidence: 99%

“…Aggregation tests can be performed either with a citrated platelet-rich plasma (cPRP) or with a washed platelet suspension. A cPRP is obtained directly after centrifugation of whole blood collected in 0.109 or 0.129 M sodium citrate tubes, while a washed platelets suspension is obtained after centrifugation of whole collected in ACD anticoagulant (acid citrate dextrose) and resuspension of the platelets in Tyrode-albumin buffer [ 46 ]. Abnormalities in platelet aggregation are often moderate in dense-granule deficiencies, and approximately 25% of patients with δ-SPD display no platelet function defect [ 47 ] when platelet function studies are performed in cPRP.…”

Section: Tools For the Diagnosis Of δ-Spdmentioning

confidence: 99%

“…In a washed platelet suspension ( Figure 2 ), one does not observe the same abnormalities as in cPRP after stimulation of platelets with ADP, whatever the concentration of agonist used. This is because ADP does not induce platelet degranulation or thromboxane A2 generation in the presence of 2 mM calcium [ 46 , 51 , 52 ] and aggregation is by definition reversible. Unlike ADP-induced aggregation which remains normal, collagen (1.25 and 2.5 µg/mL)-induced aggregation generally displays more pronounced anomalies than those observed in cPRP.…”

Section: Tools For the Diagnosis Of δ-Spdmentioning

confidence: 99%

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Platelet δ-Storage Pool Disease: An Update

Dupuis

Bordet

Eckly

et al. 2020

JCM

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Platelet dense-granules are small organelles specific to the platelet lineage that contain small molecules (calcium, adenyl nucleotides, serotonin) and are essential for the activation of blood platelets prior to their aggregation in the event of a vascular injury. Delta-storage pool diseases (δ-SPDs) are platelet pathologies leading to hemorrhagic syndromes of variable severity and related to a qualitative (content) or quantitative (numerical) deficiency in dense-granules. These pathologies appear in a syndromic or non-syndromic form. The syndromic forms (Chediak–Higashi disease, Hermansky–Pudlak syndromes), whose causative genes are known, associate immune deficiencies and/or oculocutaneous albinism with a platelet function disorder (PFD). The non-syndromic forms correspond to an isolated PFD, but the genes responsible for the pathology are not yet known. The diagnosis of these pathologies is complex and poorly standardized. It is based on orientation tests performed by light transmission aggregometry or flow cytometry, which are supplemented by complementary tests based on the quantification of platelet dense-granules by electron microscopy using the whole platelet mount technique and the direct determination of granule contents (ADP/ATP and serotonin). The objective of this review is to present the state of our knowledge concerning platelet dense-granules and the tools available for the diagnosis of different forms of δ-SPD.

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Section: Aggregation Testsmentioning

confidence: 99%

Section: Tools For the Diagnosis Of δ-Spdmentioning

confidence: 99%

“…In a washed platelet suspension ( Figure 2 ), one does not observe the same abnormalities as in cPRP after stimulation of platelets with ADP, whatever the concentration of agonist used. This is because ADP does not induce platelet degranulation or thromboxane A2 generation in the presence of 2 mM calcium [ 46 , 51 , 52 ] and aggregation is by definition reversible. Unlike ADP-induced aggregation which remains normal, collagen (1.25 and 2.5 µg/mL)-induced aggregation generally displays more pronounced anomalies than those observed in cPRP.…”

Section: Tools For the Diagnosis Of δ-Spdmentioning

confidence: 99%

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Platelet δ-Storage Pool Disease: An Update

Dupuis

Bordet

Eckly

et al. 2020

JCM

Self Cite

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“…In addition, relatively large amounts of fresh blood are required, which makes it impossible to ship samples, and requires patients to be at the place of analysis. For aggregometry, a minimum of platelet concentration (i.e., not less than 80 × 10 3 per µl) is required within platelet-rich plasma or whole blood for effective analysis [63][64][65]. This can pose a challenge when dealing with patients having moderate or severe thrombocytopenia.…”

Section: Current Diagnostic Tools For Ipdsmentioning

confidence: 99%

Diagnosis of Inherited Platelet Disorders on a Blood Smear

Zaninetti

Greinacher

2020

JCM

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Inherited platelet disorders (IPDs) are rare diseases featured by low platelet count and defective platelet function. Patients have variable bleeding diathesis and sometimes additional features that can be congenital or acquired. Identification of an IPD is desirable to avoid misdiagnosis of immune thrombocytopenia and the use of improper treatments. Diagnostic tools include platelet function studies and genetic testing. The latter can be challenging as the correlation of its outcomes with phenotype is not easy. The immune-morphological evaluation of blood smears (by light- and immunofluorescence microscopy) represents a reliable method to phenotype subjects with suspected IPD. It is relatively cheap, not excessively time-consuming and applicable to shipped samples. In some forms, it can provide a diagnosis by itself, as for MYH9-RD, or in addition to other first-line tests as aggregometry or flow cytometry. In regard to genetic testing, it can guide specific sequencing. Since only minimal amounts of blood are needed for the preparation of blood smears, it can be used to characterize thrombocytopenia in pediatric patients and even newborns further. In principle, it is based on visualizing alterations in the distribution of proteins, which result from specific genetic mutations by using monoclonal antibodies. It can be applied to identify deficiencies in membrane proteins, disturbed distribution of cytoskeletal proteins, and alpha as well as delta granules. On the other hand, mutations associated with impaired signal transduction are difficult to identify by immunofluorescence of blood smears. This review summarizes technical aspects and the main diagnostic patterns achievable by this method.

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“…This, for example, may help distinguish platelet type from plasma VWF disease. 7 This article aims to describe the different approaches used to diagnose and characterize CPDF. A description of the main results of the platelet exploration is given in ►Fig.…”

Section: Global Platelet Functional Testsmentioning

confidence: 99%

Laboratory Techniques Used to Diagnose Constitutional Platelet Dysfunction

2020

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Platelets play a major role in primary hemostasis, where activated platelets form plugs to stop hemorrhaging in response to vessel injuries. Defects in any step of the platelet activation process can cause a variety of platelet dysfunction conditions associated with bleeding. To make an accurate diagnosis, constitutional platelet dysfunction (CPDF) should be considered once von Willebrand disease and drug intake are ruled out. CPDF may be associated with thrombocytopenia or a genetic syndrome. CPDF diagnosis is complex, as no single test enables the analysis of all aspects of platelet function. Furthermore, the available tests lack standardization, and repeat tests must be performed in specialized laboratories especially for mild and moderate forms of the disease. In this review, we provide an overview of the laboratory tests used to diagnose CPDF, with a focus on light transmission platelet aggregation (LTA), flow cytometry (FC), and granules assessment. Global tests, mainly represented by LTA, are often initially performed to investigate the consequences of platelet activation on platelet aggregation in a single step. Global test results should be confirmed by additional analytical tests. FC represents an accurate, simple, and reliable test to analyze abnormalities in platelet receptors, and granule content and release. This technique may also be used to investigate platelet function by comparing resting- and activated-state platelet populations. Assessment of granule content and release also requires additional specialized analytical tests. High-throughput sequencing has become increasingly useful to diagnose CPDF. Advanced tests or external research laboratory techniques may also be beneficial in some cases.

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Platelet preparation for function testing in the laboratory and clinic: Historical and practical aspects

Cited by 46 publications

References 53 publications

Platelet δ-Storage Pool Disease: An Update

Platelet δ-Storage Pool Disease: An Update

Diagnosis of Inherited Platelet Disorders on a Blood Smear

Laboratory Techniques Used to Diagnose Constitutional Platelet Dysfunction

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