Purification of murine Kupffer cells by centrifugal elutriation.
Procedures to reproducibly obtain pure preparations of murine Kupffer cells are described. Pure Kupffer cell preparations obtained following collagenase digestion, metrizamide separation and centrifugal elutriation remained viable and maintained their phagocytic functions for at least 4-5 days in vitro. Furthermore, we determined the feasibility of extracting RNA from Kupffer cells obtained immediately following elutriation and after 2 and 4 days of culture in vitro. These RNA extracts were used to determine the level of cytokine gene expression in Kupffer cells.
Janousek
J
J
McGill Centre for the Study of Host Resistance, Montreal General Hospital Research Institute, Quebec, Canada.
Strmen
E
E
Gervais
F
F
D000818:Animals; D002469:Cell Separation / Q000379:methods; D002498:Centrifugation / Q000379:methods; D015870:Gene Expression; D007375:Interleukin-1 / Q000235:genetics; D007728:Kupffer Cells; D008099:Liver / Q000166:cytology*; D051379:Mice; D008805:Mice, Inbred A; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha