Functional activity of isolated pig hepatocytes attached to different extracellular matrix substrates. Implication for application of pig hepatocytes in a bioartificial liver

For the manufacture of a bioartifical liver for human application, large amounts of viable and active hepatocytes are needed. Pig hepatocytes are considered to be the best alternative to scarce human hepatocytes. In vitro hepatocyte functions have so far been tested under different circumstances, mainly with rat hepatocytes. Pig hepatocytes were isolated with a simple two-step isolation procedure, resulting in a high yield of viable hepatocytes. The hepatocytes were tested for their ability to synthesise urea, to metabolise 7-ethoxycoumarin (cytochrome P450 activity), and to synthesise and secrete proteins. These activities of hepatocytes while attached to tissue culture plastic were compared to the activity of the cells attached to several extracellular matrix constituents: collagen I and IV, laminim, fibronectin, Engelbreth-Holm-Swarm Natrix and in the presence of Matrigel. With the exception of Matrigel, neither of the extra-cellular matrix substrates enhanced pig hepatocyte function compared to tissue culture plastic. However, relatively large amounts of murine proteins leak out of the Matrigel. The advisability of using Matrigel or other extracellular matrix proteins in a bioartificial liver loaded with pig hepatocytes is discussed.