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Differentiation of human pluripotent stem cells to brain microvascular endothelial cell-like cells suitable to study immune cell interactions

Hideaki Nishihara, Benjamin D. Gastfriend, Pelin Kasap, Sean P. Palecek, Eric V. Shusta, Britta Engelhardt

2021STAR Protocols50 citationsDOIOpen Access PDF

Abstract

We describe the extended endothelial cell culture method (EECM) for the differentiation of human pluripotent stem cells (hPSCs) into brain microvascular endothelial cell (BMEC)-like cells. EECM-BMEC-like cells resemble primary human BMECs in morphology, molecular junctional architecture, and diffusion barrier characteristics. A mature immune phenotype with proper endothelial adhesion molecule expression makes this model distinct from any other hPSC-derived in vitro blood-brain barrier (BBB) model and suitable to study immune cell migration across the BBB in a disease relevant and personalized fashion. For complete details on the use and execution of this protocol, please refer to Lian et al. (2014), Nishihara et al. (2020a).

Topics & Concepts

Induced pluripotent stem cellCell biologyEndothelial stem cellBiologyImmune systemCellPhenotypeCellular differentiationCell typeBlood–brain barrierStem cellIn vitroImmunologyNeuroscienceEmbryonic stem cellGeneGeneticsCentral nervous systemSingle-cell and spatial transcriptomicsBarrier Structure and Function StudiesPluripotent Stem Cells Research
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