title: High Content Imaging and Analysis Enable Quantitative In Situ Assessment of CYP3A4 Using Cryopreserved Differentiated HepaRG Cells.
abstract: High-throughput imaging-based hepatotoxicity studies capable of analyzing individual cells in situ hold enormous promise for drug safety testing but are frequently limited by a lack of sufficient metabolically competent human cells. This study examined cryopreserved HepaRG cells, a human liver cell line which differentiates into both hepatocytes and biliary epithelial cells, to determine if these cells may represent a suitable metabolically competent cellular model for novel High Content Analysis (HCA) applications. Characterization studies showed that these cells retain many features characteristic of primary human hepatocytes and display significant CYP3A4 and CYP1A2 induction, unlike the HepG2 cell line commonly utilized for HCA studies. Furthermore, this study demonstrates that CYP3A4 induction can be quantified via a simple image analysis-based method, using HepaRG cells as a model system. Additionally, data demonstrate that the hepatocyte and biliary epithelial subpopulations characteristic of HepaRG cultures can be separated during analysis simply on the basis of nuclear size measurements. Proof of concept studies with fluorescent cell function reagents indicated that further multiparametric image-based assessment is achievable with HepaRG. In summary, image-based screening of metabolically competent human hepatocyte models cells such as HepaRG offers novel approaches for hepatotoxicity assessment and improved drug screening tools.
authors.affiliation: Molecular and Cell Biology Systems, EMD Millipore Corporation, 290 Concord Road, Billerica, MA 01821, USA.
authors.forename: Aarati R
authors.initials: AR
authors.lastname: Ranade
authors.affiliation: Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, MO 63132, USA.
authors.forename: Melinda S
authors.initials: MS
authors.lastname: Wilson
authors.affiliation: Barnes-Jewish Hospital, One Barnes-Jewish Hospital Plaza, St. Louis, MO 63110, USA.
authors.forename: Amy M
authors.initials: AM
authors.lastname: McClanahan
authors.affiliation: Molecular and Cell Biology Systems, EMD Millipore Corporation, 290 Concord Road, Billerica, MA 01821, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Ball