Epithelial raft cultures for investigations of virus growth, pathogenesis and efficacy of antiviral agents.
The organotypic epithelial raft cultures, originally developed to study keratinocytes differentiation, represent a novel approach to the study of viruses able to infect epithelial cells. Organotypic epithelial raft cultures accurately reproduce the process of epithelial differentiation in vitro and can be prepared from normal keratinocytes, explanted epithelial tissue, or established cell lines. This culture system permits cells to proliferate and fully differentiate at the air-liquid interface on a dermal-equivalent support. Normal primary human keratinocytes (PHKs) stratify and fully differentiate in a manner similar to the normal squamous epithelial tissues, while transformed cell lines exhibit dysplastic morphologies similar to the (pre)neoplastic lesions seen in vivo. This three-dimensional (3D) culture system provides an essential tool for investigations of virus growth, virus-host cell interactions, for the genetic analysis of viral proteins and regulatory sequences, and for the evaluation of antiviral agents. The 3D epithelial cultures have proven a breakthrough in the research on papillomaviruses, since their life cycle is strictly linked to the differentiation of the host epithelium. In the last years, several reports have shown the usefulness of the 3D epithelial cultures for the study of other viruses that target at least during a part of their life cycles epithelial cells. The 3D epithelial cultures allow the analysis of virus-host cell interactions in stratified epithelia that more closely resemble the in vivo situation. In this review we describe the advances on research on 3D epithelial cultures for the study of virus growth and pathogenesis of different families of viruses, including papilloma-, herpes-, pox-, adeno-, and parvoviruses.
Andrei
G
G
Rega Institute for Medical Research, K.U.Leuven, Belgium. graciela.andrei@rega.kuleuven.ac.be <graciela.andrei@rega.kuleuven.ac.be>
Duraffour
S
S
Van den Oord
J
J
Snoeck
R
R
D000998:Antiviral Agents / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000821:virology*; D006801:Humans; D009924:Organ Culture Techniques / Q000379:methods; D014780:Viruses / Q000187:drug effects* / Q000472:pathogenicity*
D000998:Antiviral Agents