Functional maintenance of hepatocytes on collagen gel cultured with simple serum-free medium containing sodium selenite.

We found that simple serum-free medium containing sodium selenite (Se) is effective for long-term maintenance of functional hepatocytes cultured on a pepsin-digested collagen gel (DC-gel). The concentration of Se was important for maintenance of hepatocytes, and its optimal concentration was 10(-7) approximately 10(-6)M. The effect of Se was specific, as other metals did not have the same effect. The effect was equal to that of fetal bovine serum for maintenance of functional hepatocytes. Using this medium, we could obtained a high level of hepatocellular function including the production of albumin and transferrin, and activity of p450 throughout a long-term culture. Matrigel was almost equal to DC-gel for albumin secretion, but less effective for transferrin secretion, and P450 activity in long-term cultures. The growth of cells on DC-gel or matrigel was not observed, and cell morphology of a round-shaped form was similar on both substrata. These results indicate that serum-free medium containing Se in a DC-gel culture system provides a simple method for long-term culture of hepatocytes.