Enrichment of hepatocytes in a HepaRG culture using spatially selective photodynamic treatment

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 cells 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 trans-differentiation can occur. For that purpose, a spatial light modulation device, connected to an optical microscope, was applied to selectively destroy epithelial cells through photodynamic therapy. Cells administered aminolevulinic acid (d-ALA) and subsequently illuminated with UV (l_activation =375nm, D=22 J/cm^2 ) could be selectively killed. This method is based on automatic bright field image processing and could be successfully employed in specific cell purification of other co-culture cellular models.

BEDNARKIEWICZ Artur;  MARCELINO RODRIGUES Robim;  WHELAN Maurice; 

2010-04-19

SPIE-SOC PHOTOPTICAL INSTRUMENTATION ENGINEERS

JRC57303

1083-3668,   

https://publications.jrc.ec.europa.eu/repository/handle/JRC57303,