Infectious prions accumulate to high levels in non proliferative C2C12 myotubes.
Prion diseases are driven by the strain-specific, template-dependent transconformation of the normal cellular prion protein (PrP(C)) into a disease specific isoform PrP(Sc). Cell culture models of prion infection generally use replicating cells resulting in lower levels of prion accumulation compared to animals. Using non-replicating cells allows the accumulation of higher levels of PrP(Sc) and, thus, greater amounts of infectivity. Here, we infect non-proliferating muscle fiber myotube cultures prepared from differentiated myoblasts. We demonstrate that prion-infected myotubes generate substantial amounts of PrP(Sc) and that the level of infectivity produced in these post-mitotic cells, 10(5.5) L.D.50/mg of total protein, approaches that observed in vivo. Exposure of the myotubes to different mouse-adapted agents demonstrates strain-specific replication of infectious agents. Mouse-derived myotubes could not be infected with hamster prions suggesting that the species barrier effect is intact. We suggest that non-proliferating myotubes will be a valuable model system for generating infectious prions and for screening compounds for anti-prion activity.
Centre for Prions and Protein Folding Diseases, University of Alberta, Edmonton, Alberta, Canada ; Department of Agriculture, Food and Nutritional Sciences, University of Alberta, Edmonton, Alberta, Canada.
Allen
A
Herbst
Pamela
P
Banser
Camilo Duque
CD
Velasquez
Charles E
CE
Mays
Valerie L
VL
Sim
David
D
Westaway
Judd M
JM
Aiken
Debbie
D
McKenzie
D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D006224:Cricetinae; D051379:Mice; D018485:Muscle Fibers, Skeletal; D018620:PrPSc Proteins / Q000378:metabolism*; D013045:Species Specificity
D018620:PrPSc Proteins