Enhanced maintenance and functions of rat hepatocytes induced by combination of on-site oxygenation and coculture with fibroblasts.
In in vivo liver tissue, each hepatocyte has intimate interactions not only with adjacent hepatocytes but also with nonparenchymal cells in a three-dimensional (3D) manner. We recently reported that hepatic function is highly maintained on collagen covalently immobilized poly-dimethylsiloxane (PDMS) membranes through which oxygen is supplied directly to the cells. In this study, to further enhance performances of hepatocytes culture, we investigated cocultivation of rat hepatocytes with a mouse fibroblast, NIH/3T3 (3T3) in the same PDMS membranes. Various functions of hepatocytes were better maintained on the membrane at remarkably higher levels, particularly albumin secretion on such coculture was about 20 times higher than that in conventional coculture on tissue-culture-treated polystyrene (TCPS) surfaces. The remarkable functional enhancements are likely to be explained by the net growth of hepatocytes (from 1.2- to 1.4-fold inoculated number) and very intimate contact between hepatocytes and 3T3 cells in almost continuous double-layered structures under the adequate oxygen supply. The results demonstrate that simultaneous realization of different requirements toward mimicking in vivo liver tissue microstructure is effective in improving performance of hepatocytes culture system.
Nishikawa
Masaki
M
Institute of Industrial Science (IIS), University of Tokyo, Tokyo, Japan. masaki@iis.u-tokyo.ac.jp
Kojima
Nobuhiko
N
Komori
Kikuo
K
Yamamoto
Takatoki
T
Fujii
Teruo
T
Sakai
Yasuyuki
Y
D000418:Albumins / Q000378:metabolism; D000641:Ammonia / Q000302:isolation & purification; D000818:Animals; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods*; D003470:Culture Media; D004129:Dimethylpolysiloxanes; D004801:Eosine Yellowish-(YS); D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D006416:Hematoxylin; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D010100:Oxygen / Q000378:metabolism*; D051381:Rats; D013997:Time Factors; D014508:Urea / Q000378:metabolism
D000418:Albumins; D003470:Culture Media; D004129:Dimethylpolysiloxanes; D000641:Ammonia; D014508:Urea; D010100:Oxygen; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin