Preservation of Porcine Hepatocytes in Three-Dimensional Bioreactor at Room Temperature Using Epigallocatechin-3-Gallate

Abstract

A bioartificial liver (BAL) assist system employing a three-dimensional (3D) bioreactor has been studied as a temporary support in liver failure. In the present study, a novel preservation method of primary cultured porcine hepatocytes in monolayer and 3D culture systems was studied. Epigallocatechin-3-gallate (EGCG), which has recently been found to have various bioactivities, was selected as a key compound for hepatocyte preservation. Hepatocytes isolated from porcine liver using the collagenase perfusion method were pre-cultured for 6 days, preserved at room temperature in the presence of EGCG at various concentrations for 4 days, and post-cultured in normal medium for another 6 days. In the monolayer culture, only albumin production rate was fully recovered after preservation when EGCG concentration was high (0.25 mg/mL). In contrast, albumin production and ammonium metabolism in the 3D bioreactor under the same condition recovered to 72 ± 16% and 98 ± 32%, respectively, of levels before preservation. These results indicate that hepatocytes can be preserved in the presence of 0.25 mg/mL of EGCG at room temperature, especially in a 3D culture system, which is promising technology for BAL preparation.