Premium
Isolation of rat Kupffer cells: a combined methodology for highly purified primary cultures
Author(s) -
Valatas V.,
Xidakis C.,
Roumpaki H.,
Kolios G.,
Kouroumalis E.A.
Publication year - 2003
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/s1065-6995(02)00249-4
Subject(s) - elutriation , centrifugation , kupffer cell , nitric oxide , isolation (microbiology) , differential centrifugation , microbiology and biotechnology , immunophenotyping , chemistry , biochemistry , biology , chromatography , immunology , flow cytometry , endocrinology , organic chemistry
We report a four‐step procedure that optimizes the methodology for isolation of highly purified rat Kupffer cells (KC). We combined the previously reported techniques of enzymatic tissue treatment, density gradient centrifugation, centrifugal elutriation and selective adherence. ED‐2 immunophenotyping and non‐specific esterase histochemistry were used for cell identification. This combination resulted in a satisfactorily high yield of 80–100×10 6 KCs per liver, over 95% positive for ED‐2 and 98% viable cells. Cultures of isolated KCs were functionally intact and exhibited a concentration and time‐dependent LPS‐induced TNF‐α and nitric oxide production.