Standardized isolation and culture of murine liver sinusoidal endothelial cells

Curr Protoc Cell Biol. 2014 Dec 1:65:2.9.1-8. doi: 10.1002/0471143030.cb0209s65.

Abstract

Isolation of murine liver sinusoidal endothelial cells (LSECs) is an exacting and finicky procedure. After exhaustive standardization, we were able to devise an easily reproducible protocol which produced consistent results. Moreover, we scripted a protocol which clarifies even the smallest of steps, following which isolation of LSECs is made significantly easier. Using the standardized LSEC isolation protocol herein, we demonstrated that the bacterial toxin pyocyanin (from Pseudomonas aeruginosa) induced a significant dose-dependent reduction in LSEC porosity, this being preventable by the enzyme catalase, but not by the enzyme superoxide dismutase.

Keywords: LSECs; Pseudomonas aeruginosa; cell culture; hepatocytes; liver sinusoidal endothelial cells; portal; pyocyanin.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Cell Culture Techniques / methods*
  • Cell Culture Techniques / standards
  • Cell Separation / methods*
  • Cell Separation / standards
  • Endothelial Cells* / cytology
  • Endothelial Cells* / metabolism
  • Humans
  • Liver* / cytology
  • Liver* / metabolism
  • Mice
  • Pseudomonas aeruginosa / chemistry
  • Pyocyanine / chemistry

Substances

  • Pyocyanine