An optimized protocol to isolate quiescent washed platelets from human whole blood and generate platelet releasate under clinical conditions

Luisa Weiss; Hayley MacLeod; Shane P Comer; Sarah Cullivan; Paulina B Szklanna; Fionnuala Ní Áinle; Barry Kevane; Patricia B Maguire

doi:10.1016/j.xpro.2023.102150

An optimized protocol to isolate quiescent washed platelets from human whole blood and generate platelet releasate under clinical conditions

STAR Protoc. 2023 Apr 18;4(2):102150. doi: 10.1016/j.xpro.2023.102150. Online ahead of print.

Authors

Luisa Weiss¹, Hayley MacLeod¹, Shane P Comer¹, Sarah Cullivan², Paulina B Szklanna¹, Fionnuala Ní Áinle³, Barry Kevane⁴, Patricia B Maguire⁵

Affiliations

¹ UCD Conway SPHERE Research Group, Conway Institute, University College Dublin, Dublin, Ireland; School of Biomolecular and Biomedical Science, University College Dublin, Dublin, Ireland.
² Department of Respiratory Medicine, Mater Misericordiae University Hospital, Dublin, Ireland.
³ UCD Conway SPHERE Research Group, Conway Institute, University College Dublin, Dublin, Ireland; Department for Haematology, Mater Misericordiae University Hospital, Dublin, Ireland; School of Medicine, University College Dublin, Dublin, Ireland; Department for Haematology, Rotunda Hospital, Dublin, Ireland.
⁴ UCD Conway SPHERE Research Group, Conway Institute, University College Dublin, Dublin, Ireland; Department for Haematology, Mater Misericordiae University Hospital, Dublin, Ireland; School of Medicine, University College Dublin, Dublin, Ireland.
⁵ UCD Conway SPHERE Research Group, Conway Institute, University College Dublin, Dublin, Ireland; School of Biomolecular and Biomedical Science, University College Dublin, Dublin, Ireland; Institute for Discovery, O'Brien Centre for Science, University College Dublin, Dublin, Ireland. Electronic address: [email protected].

Abstract

The contents of the platelet releasate (PR) play significant roles in hemostasis, inflammation, and pathologic sequelae. Careful platelet isolation to ensure quiescence and subsequent activation is key to the successful generation of PR. Here, we describe steps to isolate and aggregate quiescent washed platelets from whole blood of a clinical patient cohort. We then detail the generation of PR from isolated human washed platelets under clinical conditions. This protocol allows the investigation of platelet cargoes released through various activation pathways.

Keywords: Cell Biology; Cell Separation/Fractionation; Cell isolation; Clinical Protocol; Protein Biochemistry; Proteomics.