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 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 (delta-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.
Bednarkiewicz
Artur
A
European Commission, Institute of Health and Customer Protection, Via Enrico Fermi 1, Ispra, 21020 Italy. a.bednarkiewicz@int.pan.wroc.pl
Rodrigues
Robim M
RM
Whelan
Maurice P
MP
D006528:Carcinoma, Hepatocellular / Q000473:pathology* / Q000503:physiopathology*; D002454:Cell Differentiation / Q000187:drug effects / Q000528:radiation effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects / Q000528:radiation effects; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000008:administration & dosage*
D017319:Photosensitizing Agents