Mass cytometry of platelet-rich plasma: a new approach to analyze platelet surface expression and reactivity

Abstract

Mass cytometry (CyTOF) is a new technology that allows the investigation of protein expression at single cell level with high resolution. While several protocols are available to investigate leukocyte expression, platelet staining and analysis with CyTOF have been described only from whole blood. Moreover, available protocols do not allow sample storage but require fresh samples to be obtained, processed, and measured immediately. We provide a structured and reproducible method to stain platelets from platelet-rich plasma to study thrombocyte protein expression and reactivity using mass cytometry. With our method, it is possible to acquire a large number of events allowing deep bioinformatic investigation of platelet expression heterogeneity. Integrated in our protocol is also a previously established freezing protocol that allows the storage of stained samples and to delay their measurement. Finally, we provide a structured workflow using different platelet stimulators and a freely available bioinformatic pipeline to analyze platelet expression. Our protocol unlocks the potential of CyTOF analysis for studying platelet biology in health and disease.

Keywords:

• Mass cytometry
• platelet heterogeneity
• platelet reactivity
• platelets

Acknowledgements

The authors thank the Core Facility Cell Analysis at TranslaTUM (CFCA), Klinikum rechts der Isar of Technical University Munich for their support during this study. We thank Lis Arend, Judith Bernett and Quirin Manz for excellent technical assistance in the computational analysis.

Author contributions

M.K., D.B. and I.B. designed the research and wrote the paper. M.K., K.K. and J.H. performed the experiments and made the figures. K.K., O.L., M.L. and J.B. did the bioinformatic analyses. G.V. recruited the donors. M.K. and M.R. acquired the data at the CyTOF. M.S., J.R., G.C., K.L., M.L., I.B. and D.B. interpreted the results and reviewed the paper.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by the German Center for Cardiovascular Research (DZHK grant number Deutsches Zentrum für Herz-Kreislaufforschung 81 × 3600606 to D.B.). The work of O.L. was funded by the Bavarian State Ministry of Science and the Arts as part of the Bavarian Research Institute for Digital Transformation (bidt). The work of J.H. has been supported by a grant from the China Scholarship Council (CSC).