Rapid Formation of Multicellular Spheroids of Adult Rat Hepatocytes by Rotation Culture and Their Immobilization within Calcium Alginate

摘要

Abstract:Tyrosine aminotransferase (TAT) induction and albumin secretion abilities were examined in rat hepatocytes immobilized within calcium alginate; the immobilized hepatocytes lost these abilities within a week. An attempt was then made to immobilize multicellular spheroids of hepatocytes for the purpose of stabilizing the liver functions. Although it takes at least 4 days to form spheroids in the conventional method using monolayer‐cultured cells, in this study we developed a new method for rapid spheroid formation. Isolated hepatocytes were seeded into a polystyrene dish and incubated on a rotary shaker. Hepatocytes started to aggregate after 6 h of the rotation culture, and spheroids approximately 100 μm in diameter formed within 24 h. The immobilized spheroids had higher TAT induction and albumin secretion abilities, which were maintained for a longer time, than the immobilized nonaggregated cells. Further stabilization was observed in immobilized heterospheroids formed in the presence of non‐parenchymal liver cells. This method for the rapid formation of spheroids consisting of hepatocytes and non‐parenchymal liver cells could be utilized in the construction of a bioartificial liver support