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Updated: Jul 12, 2025
Analyzing Platelet Subpopulations by Multi-color Flow Cytometry
Published on: June 10, 2025
Comparison of platelet proteomic profiles between children and adults reveals origins of functional differences
Andrei K Garzon Dasgupta1, Alexey A Martyanov1,2, Anastasia A Ignatova1,2
1Center for Theoretical Problems of Physico-Сhemical Pharmacology, Russian Academy of Sciences, 30 Srednyaya Kalitnikovskaya str., Moscow, 109029, Russia.
Insights
Pediatric platelets show distinct protein compositions, impacting function. These age-related differences in platelet protein content explain variations in mitochondrial activity and calcium signaling.
Area of Science:
- Hematology
- Proteomics
- Cell Biology
Background:
- Platelets are crucial for hemostasis.
- Significant functional differences exist between adult and pediatric platelets.
- Understanding these differences is vital for clinical applications.
Purpose of the Study:
- To investigate age-dependent variations in platelet protein composition.
- To correlate protein differences with functional platelet responses.
- To identify key molecular distinctions between pediatric, adolescent, and adult platelets.
Main Methods:
- Platelet-rich plasma preparation and washing.
- Live cell flow cytometry for functional assays.
- Proteomic analysis to quantify protein content.
Main Results:
- Pediatric platelets exhibit higher levels of proteins involved in mitochondrial activity, proteasome function, and vesicle transport.
- Pediatric platelets show reduced granule secretion and altered calcium handling (increased resting, decreased ADP-stimulated calcium).
- Observed functional changes are linked to increased mitochondrial and vesicle transport proteins.
Conclusions:
- Age-dependent differences in platelet function stem from variations in protein content.
- Distinct platelet proteomic profiles in pediatric and adolescent donors suggest unique physiological characteristics.
- Platelet assessment and interpretation may need age-specific considerations.
Background:
Platelets are blood cells responsible for the prevention of blood loss upon vessel wall disruption. It has been demonstrated that platelet functioning differs significantly between adult and pediatric donors. This study aimed to identify potential differences between the protein composition of platelets of pediatric, adolescent, and adult donors.
Methods:
Platelet functional testing was conducted with live cell flow cytometry. Using a straightforward approach to platelet washing based on the sequential platelets centrifugation-resuspension, we were able to obtain stable and robust proteomics results, which corresponded to previously published data.
Results:
We have identified that pediatric donors' platelets have increased amounts of proteins, responsible for mitochondrial activity, proteasome activity, and vesicle transport. Flow cytometry analysis of platelet intracellular signaling and functional responses revealed that platelets of the pediatric donors have diminished granule secretion and increased quiescent platelet calcium concentration and decreased calcium mobilization in response to ADP. We could explain the observed changes in calcium responses by the increased mitochondria protein content, and the changes in granule secretion could be explained by the differences in vesicle transport protein content.
Conclusions:
Therefore, we can conclude that the age-dependence of platelet functional responses originates from the difference in platelet protein content.
Impact:
Platelets of infants are known to functionally differ from the platelet of adult donors, although the longevity and persistivity of these differences are debatable. Pediatric donor platelets have enhanced amounts of mitochondrial, proteasomal, and vesicle transport proteins. Platelets of the pediatric donors had increased cytosolic calcium in the resting state, what is explained by the increased numbers of mitochondrial proteins. Infants had decreased platelet granule release, which resolved upon adolescence. Thus, platelets of the infants should be assessed differently from adult platelets. Differences in platelet proteomic contents persisted in adolescent groups, yet, no significant differences in platelet function were observed.
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