Premium
Evaluation of the metabolic capability of primary human hepatocytes in three‐dimensional cultures on microstructural plates
Author(s) -
Koyama Satoshi,
Arakawa Hiroshi,
Itoh Manabu,
Masuda Norio,
Yano Kentaro,
Kojima Hajime,
Ogihara Takuo
Publication year - 2018
Publication title -
biopharmaceutics and drug disposition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.419
H-Index - 58
eISSN - 1099-081X
pISSN - 0142-2782
DOI - 10.1002/bdd.2125
Subject(s) - hepatocyte , cyp1a2 , cyp2b6 , cytochrome p450 , midazolam , chemistry , acetaminophen , inducer , cell culture , cyp3a4 , phenobarbital , pharmacology , microbiology and biotechnology , metabolism , biology , biochemistry , in vitro , genetics , sedation , gene
The NanoCulture Plate (NCP) is a novel microstructural plate designed as a base for the three‐dimensional culture of cells/tissues. This study examined whether or not the metabolic capability of human primary hepatocytes is well maintained during culture on NCPs. The hepatocytes formed aggregates after seeding and their ATP content was well maintained during culture for 21 days. Expression of CYP1A2, 2B6, 2C9, 2C19, 2D6, 2E1 and 3A4 mRNAs was detected throughout the 21‐day culture period. Addition of CYP substrate drugs (midazolam, diclofenac, lamotrigine and acetaminophen) resulted in the formation of multiple metabolites with a corresponding decrease in the amounts of the unchanged compounds. The inducers omeprazole, phenobarbital and rifampicin increased the levels of CYP1A2, 2B6 and 3A4 mRNAs by 110‐fold, 12.5‐fold and 5.4‐fold, respectively, at day 2, compared with control human hepatocytes. CYP activities were also increased at 2 days after inducer treatment (CYP1A2, 2.2‐fold; CYP2B6, 20.6‐fold; CYP3A4, 3.3‐fold). The results indicate that the hepatocyte spheroids on NCP have detectable and inducible metabolic abilities during the 7‐day culture period.