Comparison of Four Blood Sampling Sites for Flow-Cytometric Analysis of Platelet Function

2003 ◽  
Vol 30 (3) ◽  
pp. 109-114 ◽  
Author(s):  
D. Scheinichen ◽  
H.-D. Kleine ◽  
M. Koksch ◽  
M. Brandl ◽  
J. Heine ◽  
...  
Keyword(s):  
Platelet Function ◽  
Flow Cytometric Analysis ◽  
Blood Sampling ◽  
Flow Cytometric

Flow cytometric analysis of platelet function

2002 ◽  
pp. 485-498 ◽  
Author(s):  
Alan D. Michelson ◽  
Marc R. Barnard ◽  
Lori A. Krueger ◽  
A. L. Frelinger ◽  
Mark I. Furman
Keyword(s):  
Platelet Function ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric

Murine Platelets Expressing a Polymorphism of Integrin β3 Associated with Acute Coronary Thrombosis in Humans.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3896-3896
Author(s):  
David A. Wilcox ◽  
Juan Fang ◽  
Bryon D. Johnson
Keyword(s):  
Platelet Function ◽  
Flow Cytometric Analysis ◽  
Thrombotic Complication ◽  
Homogeneous Population ◽  
Integrin Αiibβ3 ◽  
Integrin Β3 ◽  
Flow Cytometric ◽  
The Impact ◽  
Human Integrin

Abstract Inheritance of a naturally occurring polymorphism of the human integrin β3-subunit at amino acid 33 causes a change from Leu to Pro that has been implicated as a risk factor for thrombotic complication in humans. It is unclear; however, if Pro (also known as the platelet alloantigen PlA2 form of β3) can alter integrin activation to significantly affect platelet function in vivo, since some clinical studies do not corroborate the initial findings. Since PlA2 is a frequent variant of β3 expressed on 28% of platelets from Americans the impact of this genetic variation on platelet function could have widespread clinical relevance; therefore, we developed a murine model to determine if expression of this common polymorphism of human integrin β3 can increase platelet function and lead to an increased propensity for thrombosis within a homogeneous population of mice. This model eliminates issues arising from variation in the genetic make-up, physical environment, and/or lifestyles of humans and allows for in vivo analysis not possible with human subjects. To accomplish this, cDNA encoding each form of the human β3-subunit was subcloned into an HIV type-1 lentivirus-derived vector under the transcriptional control of the human αIIb gene promoter to direct synthesis of β3 specifically to the megakaryocyte lineage. Bone marrow was isolated from β3-deficient mice and transduced with β3 virions encoding either the PlA1 or PlA2 form of human β3, and then transplanted into lethally irradiated β3-deficient littermates. Flow cytometric analysis demonstrated stable expression of the hybrid murine/human αIIbβ3 integrin complex on the surface of circulating platelets. Immunoanalysis using monoclonal antibodies and human serum that react specifically with the PlA1 or PlA2 confirmed the identity of each alloantigen of human β3. The lentivirus contained a second transgene encoding a drug-selectable marker (P140K MGMT) that was used for in vivo enrichment of transduced cells in mice treated with two regimens of cytotoxic reagents, O6-BG/BCNU. Flow cytometric analysis showed that use of drug-selection resulted in increased expression of the integrin αIIbβ3 complex to equal levels on nearly 100% of platelets using either form of human β3. Similar to platelets from normal mice, platelets expressing each PlA form of β3 could be induced to form aggregates ex vivo upon treatment with a cocktail of physiological agonists of platelet activation (adenosine diphosphate, epinephrine and the thrombin receptor activating peptide). These results demonstrate the feasibility for targeting expression of altered forms of the human integrin β3-subunit to murine platelets and pave the way for future studies to examine and compare the effect of integrin αIIbβ3 structure on platelet function and thrombosis in vivo.

Prediction of haemorrhage in the early stage of acute myeloid leukaemia by flow cytometric analysis of platelet function

2005 ◽  
Vol 128 (4) ◽  
pp. 526-532 ◽  
Author(s):  
Eva Birgitte Leinoe ◽  
Marianne Hutchings Hoffmann ◽  
Erik Kjaersgaard ◽  
Joern Dalsgaard Nielsen ◽  
Olav Jonas Bergmann ◽  
...  
Keyword(s):  
Acute Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Platelet Function ◽  
Early Stage ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Acute Myeloid

scholarly journals Flow Cytometric Analysis of Platelet Function in Patients on Antiplatelet Therapy and Suspected Thrombocytopathy

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 1161-1161
Author(s):  
Dana Huskens ◽  
Mark Roest ◽  
Lisa Florin ◽  
Bas De Laat ◽  
Katrien Devreese
Keyword(s):  
Antiplatelet Therapy ◽  
Platelet Function ◽  
Assessment Tool ◽  
Light Transmission ◽  
Conflicts Of Interest ◽  
Flow Cytometric Analysis ◽  
Blood Platelet ◽  
Flow Cytometric ◽  
Healthy Donors ◽  
Bleeding Score

Abstract Background: Light transmission aggregometry (LTA) is still the most used test for the identification and diagnosis of platelet function defects (PFD). Disadvantages of LTA include its laborious nature, the large volumes of blood required and the relative insensitivity to small changes in platelet function. We investigated if the flow cytometry-based whole blood platelet activation test (WB-PACT) correlates with LTA or could be used as a complimentary test. Methods: WB-PACT quantifies αIIbβ3 activation and P-selectin expression in response to 3 agonists and VWF binding to platelets in response to ristocetin. In total, 212 patients (51 on dual antiplatelet therapy (acetylsalicylic acid and ADP receptor antagonist), 161 suspected for bleeding diathesis) were tested. For WB-PACT parameters, the 2.5th, 10th and 25th percentiles were determined in 56 healthy donors to score the patient samples. For LTA, maximal aggregation, disaggregation and prolongation of the lagtime in response to 4 agonists and ristocetin was scored. Patients with at least one parameter lower than the 10th percentile measured with WB-PACT in healthy donors or at least one value deviating from the normal range measured with LTA were diagnosed with PFD. A bleeding score (BS) was calculated with the ISTH/SSC bleeding assessment tool (Rodeghiero et al., JTH, 2010). Results: A moderate correlation between LTA versus WB-PACT was found with a R of 0.60 (Figure 1). In total, 95 patients were diagnosed with PFD by both tests (κ=0.40, Table 1) and BS were recorded for 28/95 patients. Of these 28 patients, 25% had an elevated BS (adapted according to gender and age: >5 in women, >3 in men and >2 in children) and 36% had a BS>3. BS were recorded for 30/38 and 22/24 patients who were diagnosed with PFD according to WB-PACT only and LTA only, respectively. Interestingly, 27% of patients with PFD according to WB-PACT had an elevated BS and 40% had a BS>3. For LTA, only 13% of patients with PFD had an elevated BS and 13% had a BS>3. Conclusions: Flow cytometric analysis of platelet function by WB-PACT gives additional value to LTA to determine PFD. Disclosures No relevant conflicts of interest to declare.

Flow-Cytometric Analysis of Mouse Platelet Function

2004 ◽  
pp. 255-268 ◽  
Author(s):  
Bernhard Nieswandt ◽  
Valerie Schulte ◽  
Wolfgang Bergmeier
Keyword(s):  
Platelet Function ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric

Flow cytometric analysis of micronucleated reticulocytes: Time- and dose-dependent response of known mutagens in mice, using multiple blood sampling

2005 ◽  
Vol 46 (1) ◽  
pp. 30-42 ◽  
Author(s):  
Marlies De Boeck ◽  
Bas-jan van der Leede ◽  
Freddy Van Goethem ◽  
Ann De Smedt ◽  
Margino Steemans ◽  
...  
Keyword(s):  
Flow Cytometric Analysis ◽  
Blood Sampling ◽  
Flow Cytometric ◽  
Multiple Blood ◽  
Dose Dependent

scholarly journals Validation of flow cytometric analysis of platelet function in patients with a suspected platelet function defect

2018 ◽  
Vol 16 (4) ◽  
pp. 689-698 ◽  
Author(s):  
I. van Asten ◽  
R. E. G. Schutgens ◽  
M. Baaij ◽  
J. Zandstra ◽  
M. Roest ◽  
...  
Keyword(s):  
Platelet Function ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Function Defect

Flow cytometric analysis of platelet function in stored platelet concentrates

1999 ◽  
Vol 20 (2) ◽  
pp. 129-139 ◽  
Author(s):  
Chen Wang ◽  
Meera Mody ◽  
Roslyn Herst ◽  
Graham Sher ◽  
John Freedman
Keyword(s):  
Platelet Function ◽  
Flow Cytometric Analysis ◽  
Platelet Concentrates ◽  
Flow Cytometric

Flow cytometric analysis of bovine CD4 and CD8 lymphocytes: influence of blood sampling and processing methods

1994 ◽  
Vol 57 (2) ◽  
pp. 163-171 ◽  
Author(s):  
N. Abella ◽  
F. Schelcher ◽  
M. Delverdier ◽  
D. Concordet ◽  
J.F. Valarcher ◽  
...  
Keyword(s):  
Flow Cytometric Analysis ◽  
Blood Sampling ◽  
Processing Methods ◽  
Flow Cytometric
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