Highly efficient SiRNA and gene transfer into hepatocyte-like HepaRG cells and primary human hepatocytes: new means for drug metabolism and toxicity studies.

Véronique Laurent (1) , Denise Glaise (2) , Tobias Nübel , David Gilot (3) , Anne Corlu (2) , Pascal Loyer (2)

1 GGB - Génétique, génomique fonctionnelle et biotechnologies (UMR 1078)
2 Foie, métabolismes et cancer
3 IGDR - Institut de Génétique et Développement de Rennes

The metabolically competent hepatocyte-like human HepaRG cells represent a suitable alternative in vitro cell model to human primary hepatocytes. Here, we describe the culture procedure required to expand progenitor HepaRG cells and to differentiate them into hepatocyte-like cells. Transient transfection of gene and siRNA into cultured cells, using nonviral strategies, is an invaluable technique to decipher gene functions. In this chapter, we detail transfection protocols for efficient transfer of plasmid DNA or siRNAs into proliferating progenitor or quiescent differentiated HepaRG cells as well as into primary hepatocytes.

Mots clés: Nucleofection™; Human hepatocytes; Transfection; Nucleofection™; Human hepatocytes; HepaRG cells; Drug metabolism; In vitro toxicity

Domaines: Hépatologie et Gastroentérologie

Véronique Laurent, Denise Glaise, Tobias Nübel, David Gilot, Anne Corlu, et al.. Highly efficient SiRNA and gene transfer into hepatocyte-like HepaRG cells and primary human hepatocytes: new means for drug metabolism and toxicity studies.. Methods in Molecular Biology, 2013, 987, pp.295-314. ⟨10.1007/978-1-62703-321-3_25⟩. ⟨hal-00865763⟩