Permeability of Bovine Brain Microvessel Endothelial Cells in Vitro: Barrier Tightening by a Factor Released From Astroglioma Cells

Exp Cell Res. 1992 Apr;199(2):330-40. doi: 10.1016/0014-4827(92)90442-b.

Abstract

It has been shown both in vivo and in culture that astrocytes communicate with brain microvessel endothelial cells (BMECs) to induce many of the blood-brain barrier characteristics attributed to these unique cells. However, the results using cultured cells are conflicting as to whether this communication is dependent upon cell-cell contact. In this study we used primary cultures of bovine BMECs grown as monolayers on polycarbonate filters to study the formation of the barrier in vitro and examine its modulation by rat C6 glioma cells. Effects were examined by treating postconfluent BMEC monolayers with medium conditioned continually by C6 cells from the basolateral side to mimic the in vivo orientation. Cell monolayer integrity was assessed using electrical resistance and by measuring diffusion of uncharged molecules. BMEC monolayers form a functionally polarized and leaky barrier, with maximal resistance of 160 omega . cm2 and significant flux of molecules of molecular weight less than 350 Da. Treatment with rat or human astroglioma cells rather than pericytoma cells or transformed fibroblasts results in a concentration-dependent 200-440% increase in electrical resistance and a coincident 50% decrease in permeability to sucrose and dextran (70 kDa). The decrease in passive diffusion is most likely due to a change in tight junctions and not to transcellular vesicular traffic. The findings support that astroglioma cells release one or more signals that are required for cultured BMECs to express a "differentiated" phenotype associated with a tighter barrier, increased gamma-glutamyl transpeptidase activity, and decreased pinocytic activity. The relative ease and quickness of this culture system makes it amenable to studies on cell-cell interaction and regulation of barrier maintenance.

MeSH terms

  • Animals
  • Astrocytoma / metabolism*
  • Brain / blood supply*
  • Brain / enzymology
  • Brain / physiology
  • Capillaries / cytology*
  • Capillaries / metabolism
  • Capillaries / physiology
  • Cattle
  • Cell Membrane Permeability*
  • DNA / biosynthesis
  • Electricity
  • Endothelium, Vascular / cytology*
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / physiology
  • Microscopy, Electron
  • Pinocytosis
  • gamma-Glutamyltransferase / metabolism

Substances

  • DNA
  • gamma-Glutamyltransferase