title: Long-term maintenance of liver-specific functions in cultured ES cell-derived hepatocytes with hyaluronan sponge.
abstract: This study investigated the three-dimensional culture of hepatocytes differentiated from mouse embryonic stem (ES) cells with a porous hyaluronan (HA) sponge support. Hepatocytes were immobilized within the pores of the support. Spheroids could be observed within the support, each containing between 20 and 50 hepatocytes. To examine the liver-specific functions of the hepatocytes in the culture, the levels of albumin secreted into the medium were analyzed. The secretion of albumin was stable over the course of 32 days, longer than that in both conventional monolayer and collagen sponge cultures. To elucidate further the liver-specific functions of hepatocytes embedded in the HA sponge, metabolic activities of the hepatocytes were examined for their ability to eliminate ammonia from culture media and the synthesis of urea nitrogen. While rates of ammonia removal and urea nitrogen synthesis were similar to those in both conventional monolayer and in collagen sponge cultures, these functions were maintained for longer duration in cells embedded in the HA sponge. These results demonstrate that the porous HA sponge is an effective support for the in vitro culture of ES-derived hepatocytes used for both basic and applied studies for cell transplantation.
authors.affiliation: National Cancer Center Research Institute, Chuo-ku, Tokyo, Japan.
authors.forename: Takumi
authors.initials: T
authors.lastname: Teratani
authors.forename: Gary
authors.initials: G
authors.lastname: Quinn
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Tomoya
authors.initials: T
authors.lastname: Sato
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yamanokuchi
authors.forename: Akira
authors.initials: A
authors.lastname: Asari
authors.forename: Takahiro
authors.initials: T
authors.lastname: Ochiya
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003584:Cytological Techniques / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D006820:Hyaluronic Acid; D051379:Mice; D018625:Microscopy, Atomic Force; D008858:Microscopy, Phase-Contrast; D013234:Stem Cells / Q000166:cytology
chemical_list: D049452:Green Fluorescent Proteins; D006820:Hyaluronic Acid