26] Separation and collection of rat liver macrophages

Abstract

This chapter describes all the principal liver cell types that are isolated from one rat. This protocol is improved by all the principal liver cell types to be isolated from one rat and subsequently modified to optimize the cell yield and purity for both nonparenchymal and parenchymal cell types. The chapter discusses that the isolation of Kupffer cells in high yield and purity is basically a two-step procedure: the first step is aimed at releasing these cells from the reticulum tissue in which they reside; the second step is their isolation to purity by removal of other cells and debris. The first step involves enzyme treatment; this demands a decision concerning whether the parenchymal cells are to be obtained also. If so, the enzyme cannot be pronase since it destroys the parenchymal cells. The chapter also reviews that collagenase is the enzyme of choice for isolation of the parenchymal cells; use of this enzyme does not interfere with the subsequent isolation of the nonparenchyreal cells. The second step in the procedure, the handling of the crude cell isolate requires an initial decision, this being whether the Kupffer cells are to be isolated by adhesion or by centrifugal elutriation.