Expression of CYP3A4 by an immortalized human hepatocyte line in a three-dimensional culture using a radial-flow bioreactor.
Cytochrome P450 (CYP) 3A is responsible for about 50% of drug metabolizing activity in the liver. The present study was undertaken to establish a CYP3A4-active model for in vitro analysis of human drug metabolism. The cells used were immortalized normal human fetal hepatocytes (OUMS-29) and its HNF4alpha-introduced subline (OUMS-29/H-11). The cells were cultivated under high-density three-dimensional conditions in a radial-flow bioreactor (RFB). The number of OUMS-29 cells increased 15-fold over 49 days and their apical surfaces were covered with abundant microvilli, a characteristic of hepatocytes in vivo. The amount of albumin secreted by OUMS-29 cells in the three-dimensional RFB culture was 6-fold higher than those in a monolayer culture. CYP3A4 protein and an intermediate metabolite of testosterone by CYP3A4 were detected in OUMS-29/H11 cells cultivated in RFB >29 days. These results indicate that the RFB culture of OUMS-29/H-11 cells is useful for screening and developing new drugs.
Akiyama
Ichiro
I
Department of Cell Biology, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
Tomiyama
Koji
K
Sakaguchi
Masakiyo
M
Takaishi
Mikiro
M
Mori
Masaharu
M
Hosokawa
Masakiyo
M
Nagamori
Seishi
S
Shimizu
Nobuyoshi
N
Huh
Nam-Ho
NH
Miyazaki
Masahiro
M
D000418:Albumins / Q000378:metabolism; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002453:Cell Cycle; D002460:Cell Line; D048430:Cell Shape; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005434:Flow Cytometry; D005947:Glucose / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D010100:Oxygen / Q000378:metabolism
D000418:Albumins; D003577:Cytochrome P-450 Enzyme System; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D005947:Glucose; D010100:Oxygen