Establishment of Sandwich Cultures of Primary Human Hepatocytes.

Primary hepatocytes and their adherent cultures are still considered as the golden standard in the field of liver-based in vitro modeling. However, they cope with progressive deterioration of their in vivo-like morphological and functional phenotype. Among the various strategies that are used to counteract this dedifferentiation process is the seeding and cultivation of freshly isolated or cryopreserved hepatocytes between two layers of extracellular matrix scaffolds. This so-called sandwich culture system allows to restore cell-extracellular matrix interactions and thereby to delay dedifferentiation. The practical setup of the sandwich culture system of primary hepatocytes is described in this chapter.