title: RNA-seq of human prenantal brain endothelial and mural cells
design: To isolate vascular cells from prenatal human brain tissues, we adapted a previously published protocol that was used for the adult mouse brain(Crouch and Doetsch, 2018; Crouch et al., 2015). Briefly, brain tissue was minced with a scalpel and digested with collagenase/dispase, triturated in 2% FBS in 1X PBS with DNase, and centrifuged through 22% Percoll to remove debris. The samples were stained with antibodies against CD31, CD45, and CD13 (ANPEP). After washing, samples were resuspended in HBSS buffer with DAPI to exclude dead cells.