title: Isolation and primary culture of rat Kupffer cells.
abstract: The aim of this study was to describe a reproducible method for the isolation, purification and primary culture of rat Kupffer cells. Kupffer cells were isolated following sequential pronase/collagenase digestion of the liver and enrichment of a non-parenchymal cell fraction by a single-density gradient centrifugation step using 30% metrizamide. Kupffer cells were isolated and further purified from this cell fraction by centrifugal elutriation. Kupffer cells were isolated at 1017 g at 48-110 mL/min. All Kupffer cell fractions exhibited phagocytosis of 3 microm latex beads. Kupffer cell fractions isolated at 48 and 60 mL/min were predominantly ED2 negative while later fractions (80-110 mL/min) were ED2 positive. Kupffer cells were adherent in culture after 2 h. This method for Kupffer cell isolation resulted in a yield of 80-120 x 10(6) Kupffer cells per liver.
authors.affiliation: University Department of Medicine, Fremantle Hospital, Western Australia, Australia. jolynyk@cyllene.uwa.edu.au
authors.forename: J K
authors.initials: JK
authors.lastname: Olynyk
authors.forename: S L
authors.initials: SL
authors.lastname: Clarke
mesh_terms: D000818:Animals; D015415:Biomarkers; D018929:Cell Culture Techniques / Q000379:methods*; D007124:Immunoenzyme Techniques; D007728:Kupffer Cells / Q000166:cytology; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D015415:Biomarkers