Induction of blood-brain barrier properties in immortalized bovine brain endothelial cells by astrocytic factors

Neurosci Res. 1999 Nov;35(2):155-64. doi: 10.1016/s0168-0102(99)00079-6.

Abstract

The blood-brain barrier (B-BB) protects the free passage of substances into the brain and maintains the homeostasis of the central nervous system. It is commonly accepted that astrocytes surrounding brain endothelial cells influence the B-BB formation and the exhibition of B-BB function of capillaries. To begin the in vitro study on the B-BB, it is essential to obtain a homogenous and sufficient supply of brain endothelial cells as well as astrocytes. We thus immortalized the bovine brain endothelial cell (BBEC) by transfection of the SV40 large T antigen and obtained a single clone, t-BBEC-117, which retained the brain endothelial cell phenotype. Astrocyte in co-culture was found to tighten the intercellular contacts of the immortal cells resulting in a reduced L-glucose permeability, and its conditioned medium (CM) augmented a B-BB phenotype, alkaline phosphatase (ALP) activity. Among known astrocytic factors, only fibroblast growth factor-basic (bFGF) could mimic the actions of astrocytes as measured by L-glucose permeability and ALP activity. Moreover, anti-bFGF antibody canceled 90% of ALP activation by astrocyte CM. Basic FGF, however, failed to induce other B-BB phenotypes such as the expressions of multidrug resistance (mdr) and glucose transporter (GLUT-1) genes. These data suggest that bFGF is one of the most plausible astrocytic factors to induce the B-BB properties of immortal brain endothelial cells together with some unknown factors in the astrocyte CM.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / genetics
  • Alkaline Phosphatase / metabolism
  • Animals
  • Antigens, Differentiation / biosynthesis
  • Antigens, Viral, Tumor / genetics
  • Astrocytes / cytology
  • Astrocytes / metabolism*
  • Base Sequence
  • Blood-Brain Barrier / drug effects
  • Blood-Brain Barrier / physiology*
  • Brain / blood supply
  • Brain / cytology
  • Brain / drug effects
  • Brain / metabolism*
  • Cattle
  • Cell Membrane Permeability / drug effects
  • Cell Membrane Permeability / physiology
  • Cells, Cultured
  • Coculture Techniques
  • Culture Media, Conditioned / metabolism
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism*
  • Enzyme Activation / drug effects
  • Glucose / metabolism
  • Growth Substances / metabolism*
  • Growth Substances / pharmacology
  • Molecular Sequence Data
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Homology, Amino Acid
  • Transfection

Substances

  • ATP Binding Cassette Transporter, Subfamily B, Member 1
  • Antigens, Differentiation
  • Antigens, Viral, Tumor
  • Culture Media, Conditioned
  • Growth Substances
  • Alkaline Phosphatase
  • Glucose

Associated data

  • GENBANK/AB006985