An in vitro cellular system for generation of AA-amyloid.
Different conditions for establishing a cell culture system for generation of AA-amyloid were investigated. The most effective system was based on peritoneal macrophages from CBA/J mice that had received repeated injections of Hammersten casein, with subsequent cultivation of the cells at high density, high levels of acute phase serum, and neutral pH. Staining with Congo red, thioflavin T, and anti-AA revealed amyloid-like structures associated with macrophage clusters. The structures increased in number and size from day 2 to 6 of cell cultivation. The concentration of apoSAA in the culture medium fell markedly in the amyloid-producing cell cultures, while the SAP concentration was not reduced. The described cell culture system can be useful in studies of the influence of chaperone molecules and other factors or the formation and degradation of amyloid fibrils.
Palm
M
M
Department of Medical Microbiology, Odense University, Denmark.
Nielsen
E H
EH
Svehag
S E
SE
D000209:Acute-Phase Proteins / Q000032:analysis; D000818:Animals; D001053:Apolipoproteins / Q000032:analysis; D002364:Caseins / Q000494:pharmacology; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D005260:Female; D006651:Histocytochemistry; D008099:Liver / Q000166:cytology; D008262:Macrophage Activation; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008808:Mice, Inbred CBA; D000685:Serum Amyloid A Protein / Q000032:analysis / Q000096:biosynthesis*; D000683:Serum Amyloid P-Component / Q000032:analysis; D013154:Spleen / Q000166:cytology
D000209:Acute-Phase Proteins; D001053:Apolipoproteins; D002364:Caseins; D000685:Serum Amyloid A Protein; D000683:Serum Amyloid P-Component