Perfusion-based microfluidic device for three-dimensional dynamic primary human hepatocyte cell culture in the absence of biological or synthetic matrices or coagulants

Lab Chip. 2010 Dec 21;10(24):3380-6. doi: 10.1039/c0lc00135j. Epub 2010 Nov 8.

Abstract

We describe a perfusion-based microfluidic device for three-dimensional (3D) dynamic primary human hepatocyte cell culture. The microfluidic device was used to promote and maintain 3D tissue-like cellular morphology and cell-specific functionality of primary human hepatocytes by restoring membrane polarity and hepatocyte transport function in vitro without the addition of biological or synthetic matrices or coagulants. A unique feature of our dynamic cell culture device is the creation of a microenvironment, without the addition of biological or synthetic matrices or coagulants, that promotes the 3D organization of hepatocytes into cord-like structures that exhibit functional membrane polarity as evidenced by the expression of gap junctions and the formation of an extended, functionally active, bile canalicular network.

MeSH terms

  • Adenosine Triphosphate / chemistry
  • Bile Canaliculi / cytology
  • Cell Culture Techniques / instrumentation*
  • Cell Culture Techniques / methods*
  • Cells, Cultured
  • Coagulants / chemistry
  • Equipment Design
  • Gap Junctions
  • Hepatocytes / cytology*
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Microfluidic Analytical Techniques*
  • Microscopy, Fluorescence / methods
  • Models, Biological
  • Multidrug Resistance-Associated Proteins / metabolism
  • Perfusion

Substances

  • Coagulants
  • Multidrug Resistance-Associated Proteins
  • multidrug resistance-associated protein 2
  • Adenosine Triphosphate