Secretion of albumin and induction of CYP1A2 and CYP3A4 in novel three-dimensional culture system for human hepatocytes using micro-space plate.
We evaluated a novel primary three-dimensional culture system for human hepatocytes using micro-space plates. The functional activity of human hepatocytes in primary culture was determined by measuring albumin secretion from hepatocytes to medium and measuring expression levels of albumin, CYP1A2 and CYP3A4 mRNA. Albumin secretion was higher in micro-space plates compared with traditional plates after 72 h of culture; the levels of albumin secretion from hepatocytes to medium in culture using micro-space plates after 96 h of culture were 2.7-fold higher than those in culture using traditional plates, and secretion of albumin in micro-space plate culture subsequently remained constant. Expression levels of albumin, CYP1A2 and CYP3A4 mRNA in the culture of hepatocytes were significantly higher using micro-space plates than using traditional plates. The inducibility of CYP1A2 and CYP3A4 mRNA after exposure to inducers in hepatocyte culture on micro-space plates was comparable to that in culture on traditional plates, while expression of CYP1A2 and CYP3A4 mRNA after exposure to inducers was higher on micro-space plates than on traditional plates. The present study demonstrates that a novel primary three-dimensional culture system of cryopreserved human hepatocytes using micro-space plates could be used for evaluating the induction of drug-metabolizing enzymes in humans. This in vitro method may thus be useful for screening the induction potency of new drug candidates.
Nishimura
Masuhiro
M
Research and Development Center, Otsuka Pharmaceutical Factory Inc., Naruto, Tokushima, Japan.
Hagi
Mieko
M
Ejiri
Yoko
Y
Kishimoto
Sanae
S
Horie
Toru
T
Narimatsu
Shizuo
S
Naito
Shinsaku
S
D000418:Albumins / Q000096:biosynthesis / Q000378:metabolism*; D002478:Cells, Cultured; D015925:Cryopreservation; D019388:Cytochrome P-450 CYP1A2 / Q000096:biosynthesis*; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis*; D004790:Enzyme Induction; D022781:Hepatocytes / Q000201:enzymology* / Q000378:metabolism*; D006801:Humans; D013268:Stimulation, Chemical; D046509:Tissue Culture Techniques / Q000295:instrumentation / Q000379:methods
D000418:Albumins; C504573:CYP1A2 protein, human; D019388:Cytochrome P-450 CYP1A2; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human