Perfusion culture of fetal human hepatocytes in PDMS bioreactors

Abstract

New bioreactors composed of several layers of microstructured PDMS have been fabricated for perfusion culture of mammalian cells. We previously tested the bioreactors with human hepatocarcinoma Hep G2 and mouse fibroblast 3T3-L1 cells to investigate the feasibilities of microfluidic/MEMS bioreactors in the field of tissue engineering. As a new feature of those bioreactors, cultivation of fetal human hepatocytes (FHH) was attempted, because they have high possibility to mature in vivo with preserving their normality, which is suitable for implantation studies of in vitro reconstituted liver tissue. During the perfusion culture of the PDMS bioreactors for one week, cells showed good attachment, spreading and reaching their confluence over the channels of the bioreactors. Their daily albumin production was significantly enhanced in the perfusion culture using the PDMS bioreactors when compared with that in static culture using conventional tissue-culture-treated dishes. This conclusion is encouraging toward future liver tissue engineering based on microfabrication technologies and in vitro propagation and maturation of hepatocytes progenitors.