NDLI: Development of a DsRed-expressing HepaRG cell line for real-time monitoring of hepatocyte-like cell differentiation by fluorescence imaging, with application in screening of novel geometric microstructured cell growth substrates

Content Provider	Springer Nature Link
Author	Uta, Mihaela Sima, Livia E. Hoffmann, Patrik Dinca, Valentina Branza Nichita, rica
Copyright Year	2016
Abstract	The bipotent nature of the HepaRG cell line is a unique property among human hepatoma-derived cells. Cell treatment with specific differentiation inducers results in a mixture of hepatocyte- and biliary-like cells, accompanied by upregulation of liver-specific proteins, drug metabolizing enzymes, transcription regulators, membrane receptors or innate immune response effectors. These features make the HepaRG cells a suitable and handy replacement for primary hepatocytes, to study hepatic functions in vitro. However, cell differentiation is a long, variable process, requiring special culture conditions, while the resulting mixed cell populations is usually a major drawback. This process can potentially be controlled by interface characteristics, such as substrate topography. To screen for such novel substrates, we have first developed a new HepaRG cell line, designated as HepaRGDsRed, expressing the reporter gene DsRed. The fluorescent protein was expressed in hepatocyte- and not biliary-like cells, in a differentiation dependent-manner. We have further used replicated microstructured gradients of polydimethylsiloxane (PDMS) that allow three-dimensional manipulation in vitro, to monitor HepaRGDsRed differentiation in real time. We demonstrate that this approach enables the controlled assembly of viable hepatocyte-like cells for functional studies, which can be maintained in culture without loss of differentiation. The regulated expression of the DsRed reporter proved a valuable tool not only for rapid screening of novel cell growth substrates favoring cell differentiation, but also, to enrich the hepatocyte-like cell population by fluorescence-activated cell sorting to investigate liver-specific processes in vitro.
Starting Page	1
Ending Page	9
Page Count	9
File Format	PDF
ISSN	13872176
Journal	Biomedical Microdevices
Volume Number	19
Issue Number	1
e-ISSN	15728781
Language	English
Publisher	Springer US
Publisher Date	2017-01-09
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	HepaRG Hepatocyte-like cells HBV Differentiation Microstructured PDMS Biomedical Engineering Biological and Medical Physics, Biophysics Nanotechnology Engineering Fluid Dynamics
Content Type	Text
Resource Type	Article
Subject	Nanoscience and Nanotechnology Molecular Biology Biomedical Engineering

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