Because of their peculiar physico-chemical properties, alginate beads have often been proposed as an alternative cell immobilization matrix for many biotechnological applications. For entrapped hepatocytes perfused in a bioreactor, alginate beads have been demonstrated to promote viability and three-dimensional cell organization. In order to optimise the hepatocyte cell culture, we investigated the relationship between alginate beads properties, at high and low content of guluronic acid (G), and the relative cell viability and reorganization when perfused in a bioreactor. The primary structure of alginates did not apparently influence the hepatocytes culture in 8 h of perfusion in a bioreactor. However, our results confirm a preference for beads with a high content of G due to their superior mechanical resistance.