Prion propagation in cell culture

Abstract

During the past two decades, considerable efforts have been made to set up cellular cultures supporting the replication of prions, the infectious agents responsible of transmissible spongiform encephalopathies. As a matter of fact, prion-infected cell lines are very valuable to investigate the cell biology of both the normal and the pathological isoform of the prion protein or to develop and screen new therapeutics. In this chapter, we present a detailed protocol for the generation of prion-infected cells. We also give step-by-step procedures to test the biochemical properties (mainly protease resistance and insolubility) of abnormal PrP molecules, which detection represents a biochemical marker of prion propagation.