Session
Session 3
Board Number
42
Establishing a Retinal Pericyte Single Cell Isolation Protocol
The inner blood-retina barrier (BRB) is an essential part of maintaining homeostasis in the retina. It is made up of tightly packed endothelial cells and pericytes which are anchored in the basement membrane and covered by astrocyte endfeet. Pericytes are essential to proper functioning of the BRB. The goal of this study was to create an efficient single-cell isolation protocol where pericytes are enriched. This is a protocol that can be used for future scRNAseq studies of the retinal pericyte population and for increasing the number of pericytes in scRNAseq experiments. Through this study, an efficient retinal single-cell pericyte isolation protocol was established by enriching pericytes using antibody-coated beads and monitoring the purity of the isolated cells using as starting material retina from mice carrying a genetically encoded fluorescent tag in pericytes. This is an important procedure to be able to optimize because separating pericytes and endothelial cells by itself is difficult, and adding in enriching them makes it an even more difficult procedure.