Methodological considerations for the enrichment of bone marrow endothelial and mesenchymal stromal cells

Evan L. Dray, Carey G. Ousley, Daniel B. McKim

Research output: Contribution to journalArticlepeer-review

Abstract

Stromal cells are critical regulators of bone marrow hematopoietic niches, but assessment of their regulatory roles has been impeded by difficult and ineffective dissociation methods. Here, we methodically address bone marrow stromal cell dissociation. Yield of bone marrow CD45/Ter119/CD31+/CD202b+ endothelial cells (ECs) and CD45/Ter119/CD44/PDGFR+ mesenchymal stromal cells (MSCs) were determined by flow cytometry. Liberase DL, Collagenase D, and Dispase II (all supplemented with DNase) enhanced EC and MSC yields, with Dispase II + DNase proving most effective. Combinations of these enzymes did not exhibit additive benefits, nor did the addition of Elastase, TrypLE, Hyaluronidase, or Accutase. Similarly, common mechanical dissociation approaches also proved ineffective. However, the combination of gentle Dispase II + DNase dissociation with magnetic sorting dramatically enriched both ECs and MSCs. This work methodically addressed common approaches for bone marrow stromal dissociation and established an effective approach for enrichment.

Original languageEnglish (US)
Pages (from-to)127-136
Number of pages10
JournalMolecular Immunology
Volume131
DOIs
StatePublished - Mar 2021

Keywords

  • Bone marrow
  • Endothelial cell
  • Flow cytometry
  • Mesenchymal stromal cell
  • Stromal cell

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

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