A method for rapid flow-cytometric isolation of endothelial nuclei and RNA from archived frozen brain tissue.

ABSTRACT: Endothelial cells are important contributors to brain development, physiology, and disease. Although RNA sequencing has contributed to the understanding of brain endothelial cell diversity, bulk analysis and single-cell approaches have relied on fresh tissue digestion protocols for the isolation of single endothelial cells and flow cytometry-based sorting on surface markers or transgene expression. These approaches are limited in the analysis of the endothelium in human brain tissues, where fresh samples are difficult to obtain. Here, we developed an approach to examine endothelial RNA expression by using an endothelial-specific marker to isolate nuclei from abundant archived frozen brain tissues. We show that this approach rapidly and reliably extracts endothelial nuclei from frozen mouse

SUBMITTER: Kimble AL 

PROVIDER: S-EPMC8784313 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature