A stable and reproducible human blood-brain barrier model derived from hematopoietic stem cells

PLoS One. 2014 Jun 17;9(6):e99733. doi: 10.1371/journal.pone.0099733. eCollection 2014.

Abstract

The human blood brain barrier (BBB) is a selective barrier formed by human brain endothelial cells (hBECs), which is important to ensure adequate neuronal function and protect the central nervous system (CNS) from disease. The development of human in vitro BBB models is thus of utmost importance for drug discovery programs related to CNS diseases. Here, we describe a method to generate a human BBB model using cord blood-derived hematopoietic stem cells. The cells were initially differentiated into ECs followed by the induction of BBB properties by co-culture with pericytes. The brain-like endothelial cells (BLECs) express tight junctions and transporters typically observed in brain endothelium and maintain expression of most in vivo BBB properties for at least 20 days. The model is very reproducible since it can be generated from stem cells isolated from different donors and in different laboratories, and could be used to predict CNS distribution of compounds in human. Finally, we provide evidence that Wnt/β-catenin signaling pathway mediates in part the BBB inductive properties of pericytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / metabolism
  • Blood-Brain Barrier / cytology*
  • Capillary Permeability
  • Cell Adhesion Molecules / metabolism
  • Cell Differentiation
  • Cells, Cultured
  • Coculture Techniques
  • Endothelial Cells / metabolism
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism
  • Gene Expression
  • Hedgehog Proteins / genetics
  • Hedgehog Proteins / metabolism
  • Hematopoietic Stem Cells / physiology*
  • Humans
  • Models, Biological
  • Pericytes / physiology
  • Reproducibility of Results
  • Wnt Signaling Pathway

Substances

  • Biomarkers
  • Cell Adhesion Molecules
  • Hedgehog Proteins
  • SHH protein, human

Grant support

This work was supported by a Marie Curie-Reintegration Grant (FP7-People-2007-4-3-IRG; contract no 230929), funds of FEDER through the “Programa Operacional Factores de Competitividade- Compete” and Portuguese funds through FCT-Science and Technology Foundation (PTDC/CTM/099659/2008, PEst-C/SAU/LA0001/2013–2014; and SFRH/BD/42871/2008, a fellowship to S.A.), COMPETE funding (Project “Stem cell based platforms for Regenerative and Therapeutic Medicine”, Centro-07-ST24-FEDER-002008), FP7 (contracts 201024 and 202213 (European Stroke Network)) and PRIM (from the region Nord-Pas de Calais (France)). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.