title: A new culture technique for hepatocyte organoid formation and long-term maintenance of liver-specific functions.
abstract: To develop a useful hybrid artificial liver, it is important to use cultured hepatocytes that maintain liver-specific functions for a long time. These requirements were achieved recently by the use of a hepatocyte multicellular aggregate (organoid) with a tissue-like structure. In this study, we developed a three-dimensional culture of hepatocytes that formed an organoid. Primary rat hepatocytes were immobilized inside hollow fibers (for plasma separation) by centrifugation. Hepatocytes formed a cylindrical organoid (cylindroid) of 200 mum in diameter by day 2 of culture. We used two types of culture media, medium A (Williams' medium E containing insulin and epidermal growth factor) and medium B (Dulbecco's modified Eagle's medium containing insulin, epidermal growth factor, and hydrocortisone). In medium A, the hepatocyte cylindroid diminished after 14 days of culture and liver-specific functions of the hepatocyte cylindroid nearly disappeared after 1 month of culture. In contrast, hepatocyte cylindroid cultured in medium B maintained its morphology and liver-specific functions for 2-5 months. These results indicate that a combination of the new culture technique and suitable culture medium is effective for expression and maintenance of liver-specific functions of hepatocytes. This culture technique will be helpful in the development of a hybrid artificial liver.
authors.affiliation: Department of Chemical Engineering, Faculty of Engineering, Kyushu University, Fukuoka, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Mizumoto
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Ishihara
authors.forename: Kohji
authors.initials: K
authors.lastname: Nakazawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ijima
authors.forename: Kazumori
authors.initials: K
authors.lastname: Funatsu
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Kajiwara
mesh_terms: D000818:Animals; D001187:Artificial Organs; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D003470:Culture Media / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000502:physiology*; D017093:Liver Failure / Q000628:therapy; D008297:Male; D009924:Organ Culture Techniques; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D003470:Culture Media