NR AXSW
AU Moonen,P.; Geurts,Y.; Ruiter,S.; Bossers,A.
TI Prion Propagation in Decelerated Cell Culture
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Natural and Experimental Strains P02.27
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Currently, several distinct stable cell lines have been developed by transfection that express different types (like bovine and several ovine allelic variants) of the cellular form of the prion protein (PrPc). Those cell lines allow in vitro infection with a small number of fast replicating strains of natural sheep scrapie, and are invaluable to investigate the early molecular stages of the infection process. In order to enhance the use of in vitro cell culture, possibly even as a tool to differentiate between different strains of TSEs, it is necessary that a wider range of isolates can infect those cells (including slower replicating variants).
The reason why the infection is restricted to a few strains is still not known, but may be due to the speed of PrPsc production. If the production rate is too low, the infected cells may be outgrown by non-infected cells because they are passaged before the actual infection has been allowed to spread to a sufficient number of new cells (dilution).
To address this hypothesis we slowed or paused the growth of our available cell lines by different methods (such as decreasing the culture temperature, the serum content in the medium, and amount of nutritients in the medium). We infected these slowly propagating cells with either a fast replicating French scrapie isolate, or slower Dutch scrapie isolates. The increase in the amount of PrPsc was determined by Western blotting cell lysates at different times after infection. Preliminary data will be presented here.
AD P. Moonen, Y. Geurts, A. Bossers, Central Institute for Animal Disease Control, Bacteriology and TSEs, Netherlands; S. Ruiter, Animal Sciences Group Lelystad, Netherlands
SP englisch
PO Schottland