High and low permeability of human pluripotent stem cell–derived blood–brain barrier models depend on epithelial or endothelial features

authors

• Girard Stéphane D
• Julien-Gau Ingrid
• Molino Yves
• Combes Benjamin F
• Greetham Louise
• Khrestchatisky Michel
• Nivet Emmanuel

keywords

• BSA bovine serum albumin
• CNS central nervous system
• CspA cyclosporin A
• EC endothelial cell
• ECEM endothelial cell expansion medium
• ECIM endothelial cell induction medium
• ECM extracelullar matrix
• EpC epithelial cell
• FPKM fragment per kilobase million
• GSK3β glycogen synthase kinase 3 beta
• HBMVECs human brain microvascular endothelial cells
• HiPSCs human-induced pluripotent stem cells
• HUVECs human umbilical vein endothelial cells
• LY lucifer yellow
• MACS magnetic activated cell sorting
• NVU neurovascular unit
• Pe permeability
• Pr-1 protocol 1
• PSCs pluripotent stem cells
• RA retinoic acid
• RPKM read per kilobase million
• TEER trans-endothelial electrical resistance
• TGFβ transforming growth factor beta
• TNF-α tumor necrosis factor-alpha
• Ac-LDL acetylated low-density lipoprotein BBB blood-brain barrier bFGF basic fibroblast growth factor BMP4 bone morphogenic protein 4 BMVEC brain microvascular endothelial cells BSA bovine serum albumin CNS central nervous system CspA cyclosporin A EC endothelial cell ECEM endothelial cell expansion medium ECIM endothelial cell induction medium ECM extracelullar matrix EpC epithelial cell FPKM fragment per kilobase million GSK3β glycogen synthase kinase 3 beta HBMVECs human brain microvascular endothelial cells hiPSCs human-induced pluripotent stem cells HUVECs human umbilical vein endothelial cells LY lucifer yellow MACS magnetic activated cell sorting NVU neurovascular unit Pe permeability Pr-1 protocol 1 PSCs pluripotent stem cells RA retinoic acid RPKM read per kilobase million TEER trans-endothelial electrical resistance TGFβ transforming growth factor beta TNF-α tumor necrosis factor-alpha UM unconditioned medium VEGF vascular endothelial growth factor
• UM unconditioned medium
• Ac-LDL acetylated low-density lipoprotein
• VEGF vascular endothelial growth factor
• BBB blood-brain barrier
• BFGF basic fibroblast growth factor
• BMP4 bone morphogenic protein 4
• BMVEC brain microvascular endothelial cells
• BSA bovine serum albumin
• BSA bovine serum albumin
• BSA bovine serum albumin

abstract

Abstract The search for reliable human blood–brain barrier (BBB) models represents a challenge for the development/testing of strategies aiming to enhance brain delivery of drugs. Human‐induced pluripotent stem cells (hiPSCs) have raised hopes in the development of predictive BBB models. Differentiating strategies are thus required to generate endothelial cells (ECs), a major component of the BBB. Several hiPSC‐based protocols have reported the generation of in vitro models with significant differences in barrier properties. We studied in depth the properties of iPSCs byproducts from two protocols that have been established to yield these in vitro barrier models. Our analysis/study reveals that iPSCs derivatives endowed with EC features yield high permeability models while the cells that exhibit outstanding barrier properties show principally epithelial cell‐like (EpC) features. We found that models containing EpC‐like cells express tight junction proteins, transporters/efflux pumps and display a high functional tightness with very low permeability, which are features commonly shared between BBB and epithelial barriers. Our study demonstrates that hiPSC‐based BBB models need extensive characterization beforehand and that a reliable human BBB model containing EC‐like cells and displaying low permeability is still needed.

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