Primary Culture of Rat Hepatocytes on an Oxygen Permeable Membrane

Abstract

Primary culture of hepatocyte is usually performed in type I collagen coated dish as a monolayer culture. In usual, culture dish was stationarily incubated in a 5% CO₂: 20% O₂ atmosphere. Under such culture condition, medium layer over the cells becomes resistance of oxygen transport. Oxygen is an important factor for hepatocytes to maintain viability and liver function (1,2). It has been reported that medium depth should be not greater than 0.34 mm to maintain a monolayer of hepatocytes in the dish (3). However, the depth of culture medium becomes over 1mm for reasons such as prevention of drying and supply of nutrition. Therefore, monolayer culture of hepatocytes can not express the sufficient liver function due to the oxygen deficiency.