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Journal of Biomedical Optics

Enrichment of hepatocytes in a HepaRG culture using spatially selective photodynamic treatment
Author(s): Artur Bednarkiewicz; Robim M. Rodrigues; Maurice Patrick Whelan
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Paper Abstract

The human hepatoma HepaRG cell line is an in vitro cell model that is becoming an important tool in drug metabolism, hepatotoxicity, genotoxicity, and enzyme induction studies. The cells are highly proliferative during their undifferentiated state but once committed, they differentiate into two distinctly different cell types, namely, hepatocyte-like and biliary epithelial-like cells. The presence of the latter in the cell culture is considered to be a drawback of the cell model. Since the proliferating undifferentiated HepaRG cells have a bipotent character, the only way to improve the content ratio of hepatic versus biliary cells of differentiated HepaRG cells is to eradicate biliary cells in situ, in a way that free surface space does not become available and thus no transdifferentiation can occur. Spatially selective photodynamic therapy has proven to be effective for that purpose. First, all the cells were administered aminolevulinic acid (δ-ALA) to stimulate the synthesis of protoporphyrin IX (PpIX), a naturally occurring photosensitizer. Then, the biliary cells were automatically identified and outlined by bright-field image processing. Last, UV light patterns were projected onto the epithelial cells alone by a spatial light modulation device connected to an optical microscope; therefore, only these cells were destroyed by photodynamic therapy.

Paper Details

Date Published: 1 March 2010
PDF: 7 pages
J. Biomed. Opt. 15(2) 028002 doi: 10.1117/1.3369000
Published in: Journal of Biomedical Optics Volume 15, Issue 2
Show Author Affiliations
Artur Bednarkiewicz, Instytut Niskich Temperatur i Badan Strukturalnych (Poland)
Robim M. Rodrigues, European Commission Joint Research Ctr. (Italy)
Maurice Patrick Whelan, European Commission Joint Research Ctr. (Italy)


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