Differentiation of Brain Pericyte‐Like Cells from Human Pluripotent Stem Cell−Derived Neural Crest
Abstract
Brain pericytes regulate diverse aspects of neurovascular development and function, including blood‐brain barrier (BBB) induction and maintenance. Primary brain pericytes have been widely employed in coculture‐based in vitro models of the BBB, and a method to generate brain pericytes from human pluripotent stem cells (hPSCs) could provide a renewable, genetically tractable source of cells for BBB modeling and studying pericyte roles in development and disease. Here, we describe a protocol to differentiate hPSCs to NG2+PDGFRβ+αSMAlow brain pericyte‐like cells in 22‐25 days through a p75‐NGFR+HNK‐1+ neural crest intermediate, which mimics the developmental origin of forebrain pericytes. The resulting brain pericyte‐like cells have molecular and functional attributes of brain pericytes. We also provide protocols for maintenance, cryopreservation, and recovery of the neural crest intermediate, and for molecular and functional characterization of the resulting cells. © 2021 Wiley Periodicals LLC.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jan 01, 2021
- Source ID
- 10.1002/cpz1.21
Entities
People
- Benjamin D. Gastfriend
- Eric V Shusta
- Feifan Du
- Matthew J Stebbins
- Sean P Palecek
Organizations
- Defense Threat Reduction Agency
- National Science Foundation
- University of Wisconsin–Madison