Co-culture based blood-brain barrier in vitro model, a tissue engineering approach using immortalized cell lines for drug transport study

Appl Biochem Biotechnol. 2011 Jan;163(2):278-95. doi: 10.1007/s12010-010-9037-6. Epub 2010 Jul 24.


This study evaluated the feasibility of using commercially available immortalized cell lines in building an in vitro blood-brain barrier (BBB) co-culture model for preliminary drug development studies. Astrocytes-derived acellular extracellular matrix (aECM) was introduced in the co-culture model to provide a novel biomimetic basement membrane for the endothelial cells to form tight junctions. Trans-Endothelial Electrical Resistance (TEER) and solute mass transport studies quantitatively evaluated the tight junction formation. Immuno-fluorescence microscopy and Western blot analysis qualitatively verified the expression of occludin, one of the tight junction proteins on the samples. Experimental data from a total of 13 experiments conclusively showed that the novel BBB in vitro co-culture model with aECM (CO + aECM) is promising in terms of establishing tight junction formation represented by TEER values, transport profiles, and tight junction protein expression when compared with traditional co-culture (CO) model setup or the endothelial cells cultured alone (EC). In vitro colorimetric sulforhodamine B (SRB) assay also revealed that the "CO + aECM" samples resulted in less cell loss on the basal sides of the insert membranes than traditional co-culture models. Our novel approach using immortalized cell lines with the addition of aECM was proven to be a feasible and repeatable alternative to the traditional BBB in vitro modeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Astrocytes / chemistry
  • Biological Transport
  • Blood-Brain Barrier / metabolism
  • Blotting, Western
  • Caffeine / metabolism
  • Cell Line
  • Coculture Techniques / methods
  • Electric Impedance
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism*
  • Extracellular Matrix / metabolism
  • Fluoresceins / metabolism
  • Gene Expression
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mice
  • Models, Biological
  • Occludin
  • Rats
  • Reproducibility of Results
  • Tight Junctions / metabolism*
  • Tissue Engineering / methods


  • Fluoresceins
  • Membrane Proteins
  • Occludin
  • Ocln protein, mouse
  • Ocln protein, rat
  • Caffeine
  • fluorexon