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

• Published in: Lab on a chip 2010-01, Vol.10 (24), p.3380-3386
• Main Authors: Goral, Vasiliy N, Hsieh, Yi-Cheng, Petzold, Odessa N, Clark, Jeffery S, Yuen, Po Ki, Faris, Ronald A
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - chemistry; Bile; 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
• ISSN: 1473-0197; 1473-0189
• DOI: 10.1039/c0lc00135j

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.