Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles

STAR Protoc. 2025 Jun 20;6(2):103778. doi: 10.1016/j.xpro.2025.103778. Epub 2025 Apr 17.

Abstract

Hematopoietic stem cells (HSCs) maintain their self-renewal capacity in an autocrine manner through hematopoietic stem and progenitor cell (HSPC)-derived extracellular vesicles (EVs). Here, we present a protocol for the isolation and characterization of EVs from HSPCs starting from an in vivo murine model. We describe steps for murine bone marrow isolation, HSPC staining and sorting, HSPC-derived EV isolation, size and concentration characterization, and EV visualization and marker description. For complete details on the use and execution of this protocol, please refer to Bonora et al.1.

Keywords: Cell Biology; Cell isolation; Flow Cytometry; Metabolism; Stem Cells.

MeSH terms

  • Animals
  • Cell Separation* / methods
  • Extracellular Vesicles* / chemistry
  • Extracellular Vesicles* / metabolism
  • Flow Cytometry / methods
  • Hematopoietic Stem Cells* / cytology
  • Hematopoietic Stem Cells* / metabolism
  • Mice