Hepatocyte suspensions and cultures as tools in experimental carcinogenesis

Abstract

Isolation of preneoplastic cell populations would greatly facilitate analysis of the development of liver carcinogenesis. Suspensions of intact single cells can be prepared in an almost quantitative yield by two-step perfusion of the isolated liver. In the first step the liver is perfused with a Ca2+-free buffer (or with EGTA) in order to irreversibly cleave the desmosomes; in the second step perfusion with Ca2+-activated collagenase dissolves the collagenous extracellular matrix. The resulting single-cell suspension will be a mixture of intact normal and preneoplastic hepatocytes, other liver cell types (mostly Kupffer and endothelial cells), damaged cells, and subcellular debris. Intact hepatocytes can be purified--e.g., by differential centrifugation--but separation of preneoplastic from normal cells has not yet been achieved. Density gradient separation or selection in culture on the basis of the unique properties of preneoplastic hepatocytes (e.g., drug resistance) may prove useful. The use of hepatocyte cultures and liver-derived epithelial cell lines as test systems and models for chemical carcinogenesis in vitro is briefly reviewed.