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Impact of different tissue dissociation protocols on endothelial cell recovery from developing lungs.

ABSTRACT: Developing mouse lungs (day 14 of post-natal life) were dissociated by two different enzymatic methods, one employing commercially available Dispase, and another employing a commercial preparation of purified collagenase. The endothelial cell populations recovered from both approaches were characterized first by flow cytometry, to assess endothelial cell purity and viability. Subsequently, single-cell RNA-Seq was employed to characterize the constituent subpopulations of endothelial cells within each preparation. The single-cell RNA-Seq revealed that endothelial cell populations generated with Dispase exhibited reduced complexity, being enriched largely with one subpopulation of microvascular endothelial cells (gCap2) characterized by H2-Ab1 and Trf expression, whilst endothelial cell populations generated using collagenase yielded a complex endothelial cell population that included endothelial cell sub-types derived from the macro- and microvasculature, including the arterial and venous circulation, as well as the lymphatics.

ORGANISM(S): Mus musculus

PROVIDER: GSE236874 | GEO | 2024/04/30

REPOSITORIES: GEO

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