Double layered co-culture of hepatocytes and endothelial cell sheets by cell sheet engineering

摘要

Layered co-culture comprising monolayer hepatocyte and endothelial cell sheets was constructed by cell-sheet engineering. Endothelial cell sheets containing confluent cells and extracellular matrix (ECM) deposited during culture were obtained by using temperature-responsive culture dishes grafted with poly (JV-isopropylacrylamide). The endothelial cell sheet recovered by only reducing temperature was placed onto primary hepatocyte directly. Three-dimensional layered architecture was established in the co-culture. Hepatocytes showed the differentiated cell shape and extensive albumin expression under the layered system, while those cells rapidly lose their differentiated features at an early stage under conventional homotypic culture. In addition, endothelial cells also showed higher differentiated activity including acetylated-low density lipoprotein uptake in the layered system than those under homotypic culture. The Iayere3co-culture achieved by cell-sheet engineering would upregulate differentiated cellular functions by means of heterotypic cell-cell communication, and should contribute to the reconstruction of three-dimensional functional tissues in vitro.