Lucifer yellow-a robust paracellular permeability marker in a cell model of the human blood-brain barrier

Wanzhu Zhao, Linjiang Han, Younsoo Bae, Devika S. Manickam

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

The blood-brain barrier BBB consists of endothelial cells that form a barrier between the systemic circulation and the brain to prevent the exchange of non-essential ions and toxic substances. Tight junctions (TJ) effectively seal the paracellular space in the monolayers resulting in an intact barrier. This study describes a LY-based fluorescence assay that can be used to determine its apparent permeability coefficient (Papp) and in turn can be used to determine the kinetics of the formation of confluent monolayers and the resulting tight junction barrier integrity in hCMEC/ D3 monolayers. We further demonstrate an additional utility of this assay to determine TJ functional integrity in transfected cells. Our data from the LY Papp assay shows that the hCMEC/D3 cells seeded in a transwell setup effectively limit LY paracellular transport 7 days-post culture. As an additional utility of the presented assay, we also demonstrate that the DNA nanoparticle transfection does not alter LY paracellular transport in hCMEC/D3 monolayers.

Original languageEnglish
Article numbere58900
JournalJournal of Visualized Experiments
Volume2019
Issue number150
DOIs
StatePublished - Aug 2019

Bibliographical note

Publisher Copyright:
© 2019 Journal of Visualized Experiments.

Keywords

  • Bioengineering
  • Blood-brain barrier
  • DNA nanoparticles
  • HCMEC/D3
  • Issue 150
  • Lucifer yellow
  • Paracellular permeability
  • Transfection

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

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