Hepatocyte Isolation and Culture Methods

The liver plays a major role in in vivo metabolisms such as in synthesis of plasma proteins, blood glucose generation, urea formation, secretion of bile, and detoxification. Since most of these hepatic functions are performed by hepatocytes, parenchymal cells of the liver, there has been much attention to their use for cell therapeutic applications and the in vitro studies of liver physiology, drug metabolism, and hepatic toxicology [1]. However, hepatocytes are well known to dedifferentiate after isolation and lose much of their specific functions within two days under conventional culture conditions such as a monolayer on plastic surface [2]. Consequently, there have been many attempts to maintain and improve hepatocyte functions in vitro including manipulation of soluble factors, cell-cell and cell-matrix interactions. Sandwich configuration [3], in which hepatocytes are cultured between two hydrogel layers of collagen, and co-culture [4], in which hepatocytes are cultured together with other nonparenchymal cells, are the most remarkable techniques. In this paper, we first review hepatocyte isolation and culture techniques to secure a cell source and then examine hepatocyte transplantation (HTx) and bioartificial liver (BAL) studies and describe nonclinical but important uses of human hepatocytes for in vitro pharmacokinetic studies.