IngentaConnect Long-Term Maintenance of Liver-Specific Functions in Cultured ES ...

Authors: Teratani, Takumi¹; Quinn, Gary¹; Yamamoto, Yusuke²; Sato, Tomoya³; Yamanokuchi, Hiroko³; Asari, Akira³; Ochiya, Takahiro⁴

Source: Cell Transplantation, Volume 14, Number 9, 2005 , pp. 629-635(7)

Publisher: Cognizant Communication Corporation

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.

Keywords: Embryonic stem cells; Hepatocytes; Hyaluronan (HA) sponge; Three-dimensional; Spheroid

Document Type: Research article

Affiliations: 1: National Cancer Center Research Institute, 1-1, Tsukiji, 5-chome, Chuo-ku, Tokyo 104-0045, Japan 2: National Cancer Center Research Institute, 1-1, Tsukiji, 5-chome, Chuo-ku, Tokyo 104-0045, Japan, Department of Biology, School of Education, Waseda University, Tokyo 169-0051, Japan 3: Central Research Laboratories, Seikagaku Corporation, 1253 Tateno 3-chome, Higashiyamato-shi, Tokyo 207-0021, Japan 4: National Cancer Center Research Institute, 1-1, Tsukiji, 5-chome, Chuo-ku, Tokyo 104-0045, Japan,

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Long-Term Maintenance of Liver-Specific Functions in Cultured ES Cell-Derived Hepatocytes With Hyaluronan Sponge