Journal
CELL BIOLOGY INTERNATIONAL
Volume 27, Issue 1, Pages 67-73Publisher
ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD
DOI: 10.1016/S1065-6995(02)00249-4
Keywords
Kupffer cells; sinusoidal liver cells; cell isolation; cell culture
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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 x 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-alpha and nitric oxide production. (C) 2003 Elsevier Science Ltd. All rights reserved.
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