A new culture technique for hepatocyte organoid formation and long-term maintenance of liver-specific functions.
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.
Mizumoto
Hiroshi
H
Department of Chemical Engineering, Faculty of Engineering, Kyushu University, Fukuoka, Japan.
Ishihara
Kazuhisa
K
Nakazawa
Kohji
K
Ijima
Hiroyuki
H
Funatsu
Kazumori
K
Kajiwara
Toshihisa
T
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*
D003470:Culture Media