Journal
STEM CELL REPORTS
Volume 13, Issue 4, Pages 599-611Publisher
CELL PRESS
DOI: 10.1016/j.stemcr.2019.08.002
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Funding
- Portugal2020 project Stroke-Therapy [3386]
- EC project ERA chair (ERA@UC) [669088]
- FCT [PTDC/DTP-FTO/2784/2014, 02/SAICT/2017/029229, SFRH/BD/51678/2011, SFRH/BPD/79323/2011]
- Fundação para a Ciência e a Tecnologia [PTDC/DTP-FTO/2784/2014, SFRH/BD/51678/2011] Funding Source: FCT
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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 x 10(-3) cm/min, a transendothelial electrical resistance value of 60 Omega cm(2) 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.
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