Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system.
A micro-space cell culture system was recently developed in which cells such as hepatocytes can be cultured and formed into a multicellular three-dimensional (3D) architecture. In this study, we assessed the performance of HepG2 cells cultured in this micro-space cell culture system in a drug toxicity test, and evaluated the effects of micro-space culture on their hepatocyte-specific functions. The micro-space cell culture facilitated the formation of 3D HepG2 cell architecture. HepG2 cells cultured in a micro-space culture plate exhibited increased albumin secretion and enhanced mRNA expression levels of cytochrome P450 (CYP) enzyme compared to those cultured in a monolayer culture. When the cells were exposed to acetaminophen, a hepatotoxic drug, the damage to the HepG2 cells grown in micro-space culture was greater than the damage to the HepG2 cells grown in monolayer culture. In addition, human primary hepatocytes grown in micro-space culture also exhibited increased albumin secretion, enhanced CYP mRNA expression levels and increased sensitivity to acetaminophen compared to those grown in monolayer culture. These results suggest that this micro-space culture method enhances the hepatocyte-specific functions of hepatocytes, including drug-metabolizing enzyme activities, making hepatocytes grown in the micro-space culture system a useful tool for evaluating drug toxicity in vitro.
Nakamura
Kazuaki
K
Department of Pharmacology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan.
Mizutani
Reiko
R
Sanbe
Atsushi
A
Enosawa
Shin
S
Kasahara
Mureo
M
Nakagawa
Atsuko
A
Ejiri
Yoko
Y
Murayama
Norie
N
Miyamoto
Yuki
Y
Torii
Tomohiro
T
Kusakawa
Shinji
S
Yamauchi
Junji
J
Fukuda
Motohiro
M
Yamazaki
Hiroshi
H
Tanoue
Akito
A
D000082:Acetaminophen / Q000633:toxicity; D000418:Albumins / Q000096:biosynthesis; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D018675:Toxicity Tests
D000418:Albumins; D000082:Acetaminophen; D003577:Cytochrome P-450 Enzyme System