Identifcation and stablization of a novel 3D hepatocyte monolayer for hepatocyte-based applications

Abstract

An interesting morphological stage before the formation of 3D spheroids was discovered when the hepatocytes attach to the substrata as monolayer but exhibiting many characteristics of the 3D spheroids, but without the limitations of the 3D spheroids. We coined the name of b 3D hepatocyte monolayerb to indicate its monolayer morphology as well as 3D cell characteristics, which make it a promising novel hepatocyte culture model to improve the conventional 2D hepatocyte monolayer culture. Mechanistic studies revealed that the dynamic morphogenesis is mainly regulated by the balance between cell-cell interaction and cell-substratum interaction, which provided explanations for the existences of this 3D monolayer stage. Since the 3D hepatocyte monolayer is only a transient morphological stage existing less than two days, we employed two approaches to achieve short-term and longer-term stabilization of the 3D hepatocyte monolayer respectively, which are readily useful for the related hepatocyte-based applications.