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J. Biochem, 1998, Vol. 123, No. 6 1017-1023
© 1998 Japanese Biochemical Society

research-article

Efficient Induction of Hepatocyte Spheroids in a Suspension Culture Using a Water-Soluble Synthetic Polymer as an Artificial Matrix1

Keisuke Yamada, Masamichi Kamihira2, Ryuji Hamamoto and Shinji Iijima

Department of Biotechnology, Graduate School of Engineering, Nagoya University Chikusa-ku, Nagoya 464-8603

2To whom correspondence should be addressed. Phone: +81-52-789-4277, FAX: +81-52-789-3221, E-mail: kamihira{at}proc.nubio.nago-ya-u.ac.jp

The preparation of hepatocyte spheroids by adding a water-soluble synthetic polymer as an artificial matrix was performed in a cell suspension system. Cell-aggregation was promoted without cytotoxicity by adding Eudragit (a copolymer of methacrylic acid and methylmethacrylate) to the culture medium. Spheroid-like cell aggregates, whose liver functions were enhanced, were effectively formed in the presence of 0.1% Eudragit, independent of the cultural substratum. Moreover, the mass preparation of spheroids could be achieved with a high production yield by means of a suspension culture in a spinner flask. In this case, the polymer protected the cells from damage due to agitation. The spheroids induced with Eudragit expressed high liver functions, such as albumin secretion, ammonia removal, and urea synthesis. On histological observation, the spheroids showed a well-developed cell adhesion apparatus and bile canaliculi. In addition, a higher calcium ion concentration in the cells of spheroids was observed compared with in monolayer cells.

1This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan, and in part by the Ciba-Geigy Foundation (Japan) for the Promotion of Science.


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