Methodological considerations for the enrichment of bone marrow endothelial and mesenchymal stromal cells.
Mol Immunol
; 131: 127-136, 2021 03.
Article
em En
| MEDLINE
| ID: mdl-33441247
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.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Medula Óssea
/
Células da Medula Óssea
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Células Estromais
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Células Endoteliais
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Células-Tronco Mesenquimais
Limite:
Animals
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article