title: Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system.
abstract: 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.
authors.affiliation: Department of Pharmacology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Nakamura
authors.forename: Reiko
authors.initials: R
authors.lastname: Mizutani
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sanbe
authors.forename: Shin
authors.initials: S
authors.lastname: Enosawa
authors.forename: Mureo
authors.initials: M
authors.lastname: Kasahara
authors.forename: Atsuko
authors.initials: A
authors.lastname: Nakagawa
authors.forename: Yoko
authors.initials: Y
authors.lastname: Ejiri
authors.forename: Norie
authors.initials: N
authors.lastname: Murayama
authors.forename: Yuki
authors.initials: Y
authors.lastname: Miyamoto
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Torii
authors.forename: Shinji
authors.initials: S
authors.lastname: Kusakawa
authors.forename: Junji
authors.initials: J
authors.lastname: Yamauchi
authors.forename: Motohiro
authors.initials: M
authors.lastname: Fukuda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamazaki
authors.forename: Akito
authors.initials: A
authors.lastname: Tanoue
mesh_terms: 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
chemical_list: D000418:Albumins; D000082:Acetaminophen; D003577:Cytochrome P-450 Enzyme System