A New Bioartificial Liver Using Porcine Hepatocyte Spheroids in High-Cell-Density Suspension Perfusion Culture: In Vitro Performance in Synthesized Culture Medium and in 100% Human Plasma

Abstract

A prototype of a bioartificial liver (BAL) based on suspension perfusion culture of porcine hepatocyte spheroids was developed at 150 ml scale. About 2% (4 × 109 cells) of whole human liver cells was immobilized. The cell density in the bioreactor was 2.7 × 107 cells/ml, which was almost comparable to that of presently developed packed-bed-type BALs. The bioreactor was perfused with culture medium while retaining spheroids. This was done using a rotating stainless filter (pore size 50 μm). In vitro 8-h perfusion experiments utilizing both synthesized culture medium and 100% human plasma demonstrated the spheroids in the bioreactor had almost the same functions on a unit/cell basis as those in small-scale rotational culture. This indicated that the functional deterioration often associated with scaling up had been minimized. Rapid spheroid aggregation and dysfunction in specific human plasma pool must be eliminated before clinical application, although this phenomenon seemed to be inherent to porcine hepatocyte-based BALs. This prototype shows promise in meeting present clinical demands by achieving maximal metabolic activities even in the short term.