Preservation of porcine hepatocytes in three-dimensional bioreactor at room temperature using epigallocatechin-3-gallate.
A bioartificial liver (BAL) assist system employing a three-dimensional (3D) bioreactor has been studied as a temporary support in liver failure. In the present study, a novel preservation method of primary cultured porcine hepatocytes in monolayer and 3D culture systems was studied. Epigallocatechin-3-gallate (EGCG), which has recently been found to have various bioactivities, was selected as a key compound for hepatocyte preservation. Hepatocytes isolated from porcine liver using the collagenase perfusion method were pre-cultured for 6 days, preserved at room temperature in the presence of EGCG at various concentrations for 4 days, and post-cultured in normal medium for another 6 days. In the monolayer culture, only albumin production rate was fully recovered after preservation when EGCG concentration was high (0.25mg/mL). In contrast, albumin production and ammonium metabolism in the 3D bioreactor under the same condition recovered to 72+/-16% and 98+/-32%, respectively, of levels before preservation. These results indicate that hepatocytes can be preserved in the presence of 0.25mg/mL of EGCG at room temperature, especially in a 3D culture system, which is promising technology for BAL preparation.
Miskon
Azizi
A
Department of Biomedical Engineering, Advanced Medical Engineering Center, National Cardiovascular Center Research Institute, Osaka 565-8565, Japan.
Yamaoka
Tetsuji
T
Hyon
Suong-Hyu
SH
Kodama
Makoto
M
Uyama
Hiroshi
H
D000818:Animals; D019149:Bioreactors; D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000295:instrumentation*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D019164:Liver, Artificial; D013552:Swine; D013556:Swine, Miniature; D013696:Temperature; D023822:Tissue Engineering / Q000295:instrumentation*; D014021:Tissue Preservation / Q000295:instrumentation* / Q000379:methods
D002392:Catechin; C045651:epigallocatechin gallate