Chapter 4 Preparation of Isolated Rat Liver Cells

Publisher Summary This chapter discusses preparation of isolated rat liver cells. The early mechanical and chemical methods for liver-cell preparation were relatively successful in converting liver tissue to a suspension of isolated cells. The successful preparation of intact liver cells by perfusion with collagenase is technically quite difficult. The major method for preparation of nonparenchymal liver cells is based on the selective sensitivity of parenchymal cells toward proteases. By incubation of rat liver minces with pronase, most of the liver parenchyma is digested, while nonparenchymal cells remain intact and can be recovered from the incubate. Similar results have been reported with trypsin digestion of collagenase-dispersed liver minces, but pronase appears to be more effective. The most common procedure is to perfuse the liver briefly with pronase before it is minced and incubated with the enzyme. Such direct pronase methods have been used by several investigators with yields of nonparenchymal liver cells reported to be in the range 2–15 × 10 6 cells/gm liver.