The current status of primary hepatocyte culture

Abstract

Recently, there have been significant advances toward the development of culture conditions that promote proliferation of primary rodent hepatocytes. There are two major methods for the multiplication of hepatocytes in vitro: one is the use of nicotinamide, the other is the use of a nutrient-rich medium. In the medium containing a high concentration of nicotinamide and a growth factor, primary hepatocytes can proliferate well. In this culture condition small mononucleate cells, which are named small hepatocytes, appear and form colonies. Small hepatocytes have a high potential to proliferate while maintaining hepatic characteristics, and can differentiate into mature ones. On the other hand, combining the nutrient-rich medium with 2% DMSO, the proliferated hepatocytes can recover the hepatic differentiated functions and maintain them for a long time. In this review I describe the culture conditions for the proliferation and differentiation of primary hepatocytes and discuss the small hepatocytes, especially their roles in liver growth.

Selective proliferation of rat hepatocyte progenitor cells in serum-free culture

“…However, the effect of HA is not different among commercially available forms of HA derived from pig skin, bovine vitreous humor, rooster comb and Streptococcus (data not shown). To observe the appearance of SHs in serum-free culture, both nicotinamide and growth factors (either a single factor or a combination of EGF, hepatocyte growth factor and transforming growth factor-a) must be included in the medium 1 . These results suggest that ingredients contained in Ham F12 medium are important for SHs to grow in the serum-free culture, because the use of DMEM alone without FBS cannot support the growth of SHs.…”

“…m CRITICAL STEP If researchers need to grow MHs, we suggest using nicotinamide-supplemented medium 1 (ii) Although collagenase works efficiently at a slow flow rate, too slow a flow causes partial perfusion of the liver, and the yield and viability of the cells have a tendency to be bad…”

“…Small hepatocytes (SHs) are a subpopulation of hepatocytes that have high growth potential in culture 1 . Although the cells are less than half the size of mature hepatocytes (MHs), they possess hepatic characteristics 1 .…”

“…Although the cells are less than half the size of mature hepatocytes (MHs), they possess hepatic characteristics 1 . SHs can clonally proliferate to form a colony and can differentiate into MHs by interacting with hepatic non-parenchymal cells (NPCs) 2,3 or as a result of treatment with gel derived from Engelbreth-Holm-Swarm sarcoma 4 .…”

A Mathematical Model of Liver Cell Aggregation In Vitro

“…In vivo, under normal physiological conditions, around 80% of liver tissue is composed of hepatocytes (Mitaka, 1998). These epithelial cells perform most of the liver's important functions (Selden et al, 1999), and hence have received the greatest attention from tissue engineers.…”