Fig. 2

Summary of critical microvascular isolation steps. Through the microvascular isolation procedure, five critical steps greatly influence the effectiveness and yield of each cell type. Cells can be over-dissociated or digested during Step [1] which would be evidenced in Step [2]. Step [2] can be run again to ensure all vascular cells are captured. Similar risks of Step [1] exist at Step [3]. The final product of Step [3] is a mixed vascular pellet that is further separated at Step [4]. Four distinct layers with visible vascular fragments should be seen at Step [4]. The vascular layers in Step [4] are further separated at Step [5] where the top layer contains the endothelial cells and pericytes. To enrich for pericytes, plates should be uncoated. Collagen and fibronectin coatings are needed for endothelial cell attachment and proliferation. This figure was created with Biorender.com