Micro-nano scale mass transfer in the reconstructed hepatocyte structure

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Three-dimensional (3D) culture of hepatocytes is essential in order to reconstruct functional hepatic tissues in vitro. We have established the 3D stacked-up culture, a 3D culture method for the reconstruction of layered hepatocyte tissues which mimic the structure of hepatic cords. Hepatic stellate cells (HSCs) form a functional unit with endothelia and hepatocytes in the liver to play a pivotal role in heterotypic cellular communication. We confirmed that HSCs mediated the communication between hepatocytes and ECs in terms of EC morphogenesis. This tri-culture model allows us to investigate the roles of HSC_s as both facilitators and integrators of cell. cell communication between hepatocytes and ECs, and is useful for investigating heterotypic cellular communication in vitro. We also developed a 3D stacked-up culture for SHs based on a degradation of microporous poly (D, L-lactide-co-glycolide acids) (PLGA) membranes with defined geometry. The 3D stack-up culture based on the degradation of microporous PLGA membranes will enable reconstruction of multilayered hepatic tissues with highly differentiated functions in vitro. Finally we achieved the vascularization into the 3D reconstructed structure of hepatocyte by micro-fluidic device. Thereby the technology of reconstructed liver tissue has been advanced to the final application to the bio-artificial liver.