Isolation,culture and identification of the rat liver Kupffer cells

Objectives:Kupffer cell of the liver account for 80 to 90 percent of the mononuclear phagocytotic system (MPS), but the volume is only 2.1 percent of the liver. So it is quite difficult to separate and purify. Methods: In this experiment, the isolation techniques include:①Combined enzymatic digestion. ②Isopyknic gradient centrifugation with Nycodenz.③Selective attachment of the Kupffer cells. Results: With this method, about (28.68±4)×10 6/rat liver Kupffer cells were obtained, purity was 96%, and more than 95% cells were alive. The isolated and purified rat Kupffer cells retained their in vivo morphological, biological and immunological characteristics. Conclusions: A simple, effective and stable method was established for preparing Kupffer cells in high yield. Isolated and culture Kupffer cells retained their in vivo biological characteristics.