A new blood-brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes

Shinsuke Nakagawa, M. Deli, Hiroko Kawaguchi, Takeshi Shimizudani, Takanori Shimono, A. Kittel, Kunihiko Tanaka, Masami Niwa

Research output: Contribution to journalArticle

368 Citations (Scopus)

Abstract

Blood-brain barrier (BBB) characteristics are induced and maintained by cross-talk between brain microvessel endothelial cells and neighbouring elements of the neurovascular unit. While pericytes are the cells situated closest to brain endothelial cells morphologically and share a common basement membrane, they have not been used in co-culture BBB models for testing drug permeability. We have developed and characterized a new syngeneic BBB model using primary cultures of the three main cell types of cerebral microvessels. The co-culture of endothelial cells, pericytes and astrocytes mimick the anatomical situation in vivo. In the presence of both pericytes and astrocytes rat brain endothelial cells expressed enhanced levels of tight junction (TJ) proteins occludin, claudin-5 and ZO-1 with a typical localization at the cell borders. Further morphological evidence of the presence of interendothelial TJs was provided by electron microscopy. The transendothelial electrical resistance (TEER) of brain endothelial monolayers in triple co-culture, indicating the tightness of TJs reached 400 Ω cm2 on average, while the endothelial permeability coefficients (Pe) for fluorescein was in the range of 3 × 10-6 cm/s. Brain endothelial cells in the new model expressed glucose transporter-1, efflux transporters P-glycoprotein and multidrug resistance protein-1, and showed a polarized transport of rhodamine 123, a ligand for P-glycoprotein. To further characterize the model, drug permeability assays were performed using a set of 19 compounds with known in vivo BBB permeability. Good correlation (R2 = 0.89) was found between in vitro Pe values obtained from measurements on the BBB model and in vivo BBB permeability data. The new BBB model, which is the first model to incorporate pericytes in a triple co-culture setting, can be a useful tool for research on BBB physiology and pathology and to test candidate compounds for centrally acting drugs.

Original languageEnglish
Pages (from-to)253-263
Number of pages11
JournalNeurochemistry International
Volume54
Issue number3-4
DOIs
Publication statusPublished - Mar 2009

Fingerprint

Pericytes
Blood-Brain Barrier
Astrocytes
Endothelial Cells
Brain
Permeability
Coculture Techniques
P-Glycoprotein
Microvessels
Claudin-5
Pharmaceutical Preparations
Occludin
Tight Junction Proteins
Rhodamine 123
Facilitative Glucose Transport Proteins
Fluorescein
Electric Impedance
Basement Membrane
Electron Microscopy
Pathology

Keywords

  • Astrocytes
  • Blood-brain barrier
  • Brain endothelial cells
  • Co-culture
  • Drug permeability
  • In vitro BBB model (rat)
  • P-Glycoprotein
  • Pericytes
  • Tight junction
  • Transendothelial electrical resistance

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Cell Biology

Cite this

A new blood-brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes. / Nakagawa, Shinsuke; Deli, M.; Kawaguchi, Hiroko; Shimizudani, Takeshi; Shimono, Takanori; Kittel, A.; Tanaka, Kunihiko; Niwa, Masami.

In: Neurochemistry International, Vol. 54, No. 3-4, 03.2009, p. 253-263.

Research output: Contribution to journalArticle

Nakagawa, Shinsuke ; Deli, M. ; Kawaguchi, Hiroko ; Shimizudani, Takeshi ; Shimono, Takanori ; Kittel, A. ; Tanaka, Kunihiko ; Niwa, Masami. / A new blood-brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes. In: Neurochemistry International. 2009 ; Vol. 54, No. 3-4. pp. 253-263.
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