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Stem Cell Reports. 2019 Oct 8;13(4):599-611. doi: 10.1016/j.stemcr.2019.08.002. Epub 2019 Sep 5.

Derivation of Brain Capillary-like Endothelial Cells from Human Pluripotent Stem Cell-Derived Endothelial Progenitor Cells.

Author information

1
Doctoral Programme in Experimental Biology and Biomedicine (PDBEB), Portugal CNC - Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal; Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal; Faculty of Medicine, University of Coimbra, Coimbra, Portugal.
2
Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.
3
Faculté des Sciences Jean Perrin, Université d'Artois, Lens, France.
4
Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal; Faculty of Medicine, University of Coimbra, Coimbra, Portugal. Electronic address: lino@uc-biotech.pt.

Abstract

The derivation of human brain capillary endothelial cells is of utmost importance for drug discovery programs focusing on diseases of the central nervous system. Here, we describe a two-step differentiation protocol to derive brain capillary-like endothelial cells from human pluripotent stem cells. The cells were initially differentiated into endothelial progenitor cells followed by specification into a brain capillary-like endothelial cell phenotype using a protocol that combined the induction, in a time-dependent manner, of VEGF, Wnt3a, and retinoic acid signaling pathways and the use of fibronectin as the extracellular matrix. The brain capillary-like endothelial cells displayed a permeability to lucifer yellow of 1 × 10-3 cm/min, a transendothelial electrical resistance value of 60 Ω cm2 and were able to generate a continuous monolayer of cells expressing ZO-1 and CLAUDIN-5 but moderate expression of P-glycoprotein. Further maturation of these cells required coculture with pericytes. The study presented here opens a new approach for the study of soluble and non-soluble factors in the specification of endothelial progenitor cells into brain capillary-like endothelial cells.

KEYWORDS:

blood-brain barrier; brain capillary-like endothelial cells; extracellular matrices; induced pluripotent stem cells; in vitro model; soluble factors

PMID:
31495714
DOI:
10.1016/j.stemcr.2019.08.002
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