Drug metabolism in 3-D perfused microreactor liver cultures

TOXI 43

Ju Liu, juliu@mit.edu1, Walker Inman, winman@MIT.EDU1, Sharon Karakattu, sliz@mit.edu1, Michal Bokayza1, Karel Domansky, domansky@mit.edu2, Keith Hoffmaster, Keith.A.Hoffmaster@pfizer.com3, Linda G. Griffith, griff@mit.edu4, and Steve R. Tannenbaum, srt@mit.edu5. (1) Biological Engineering Division, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139, (2) Biotechnology Process Engineering Center, Massachusetts Institute of Technology, Room 16-436, 77 Massachusetts Avenue, Cambridge, MA 02139, (3) Pfizer Research Technology Center, 620 Memorial Dr, Cambridge, MA 02139, (4) Department of Chemical and Biological Engineering, Center for Biomedical Engineering, and Biotechnology Process Engineering Cent, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Room 66-466, Cambridge, MA 02139, (5) Division of Biological Engineering, Massachusetts Institute of Technology, 56-731, 77 Massachusetts avenue, Cambridge, MA 02139
We have developed a multi-well plate bioreactor system that fosters 3D organization of liver cells and provides local microscale perfusion. To test whether isolated rat primary hepatocytes or human cryopreserved hepatocytes in the bioreactors exhibit physiological metabolism, probe substrates for CYP 3A4 (midazolam), 1A2 (phenacetin and propranolol) and 2C9 (tolbutamide) were used in this study. A rapid quantitative LC/MS assay using single quadrupole mass spectrometer combined with UPLC chromatography was developed to analyze substrates and products. A model of flow and reaction in the system was used to extract kinetic parameters from 3D cultures. Using this method the Cl-intrinsic was estimated for the probe substrates in isolated hepatocytes and 3D microperfused culture. The data indicate that this model may be useful in evaluation of liver toxicity and drug development.
 

Poster Session and Awards
6:00 PM-10:00 PM, Tuesday, August 21, 2007 BCEC -- 204 A/B, Poster

Sci-Mix
8:00 PM-10:00 PM, Monday, August 20, 2007 BCEC -- Exhibit Hall - B2, Sci-Mix

Division of Chemical Toxicology

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007