Perfusion culture of hepatocytes within galactose-derivatized biodegradable poly(lactide-co-glycolide) scaffolds prepared by gas foaming of effervescent salts.

Abstract

Galactose, a specific ligand for asialoglycoprotein receptor in hepatocytes, was immobilized onto the internal surface of highly porous biodegradable poly(D,L-lactic-co-glycolic acid) scaffolds prepared by gas foaming of effervescent salts. Rat hepatocytes seeded within the scaffolds were cultivated by using a continuous flow and perfusion reactor system. Flow rate of medium circulating through the closed loop bioreactor system was optimized to minimize the extent of cell washout from the scaffold/cell construct while satisfying the oxygen transport rate to the seeded hepatocytes. Using the flow culture system, the scaffolds immobilized with galactose onto its internal surface retained a greater number of hepatocytes than those with unmodified or immobilized with glucose due to specific interactions between seeded hepatocytes and galactose moieties exposed onto the surface of the scaffolds. The perfusion culture system based on galactose-modified macroporous scaffolds, under optimal flow conditions, resulted in much higher albumin secretion rate, approximately 70 pg/cell/day for 7 days, compared to that with glucose modified scaffolds used as a negative control. The enhanced functional activity of hepatocytes seeded within the galactose modified scaffolds was likely caused by the formation of aggregated hepatocytes within the scaffolds.