Microfluidic-based measurements of cytochrome P450 enzyme activity of primary mammalian hepatocytes

被引：8

作者：

Anderson, Keith ^{[2
]}

Cooper, Jonathan M. ^{[3
]}

Haswell, Stephen J. ^{[1
]}

Marshall, Damian ^{[2
]}

Yin, Huabing ^{[3
]}

Zhang, Xunli ^{[1
,4
]}

机构：

[1] Univ Hull, Dept Chem, Kingston Upon Hull HU6 7RX, N Humberside, England

[2] LGC Ltd, Teddington TW11 0LY, Middx, England

[3] Univ Glasgow, Dept Elect & Elect Engn, Glasgow G12 8QQ, Lanark, Scotland

[4] Univ Southampton, Sch Engn Sci, Bioengn Grp, Southampton SO17 1BJ, Hants, England

来源：

ANALYST | 2010年 / 135卷 / 06期

关键词：

CELLS; ASSAY; METABOLISM; PROTEIN; DEVICE; CYP1A1; EROD;

D O I：

10.1039/c0an00031k

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A microfluidic-based system was developed for the in situ monitoring of the 7-ethoxyresorufin O-dealkylation (EROD) activity of primary rat hepatocytes by measuring the fluorescent intensity of both cells and their surrounding media. The microfluidic chip was designed to allow the cell suspension and test reagent to be introduced in a layer-by-layer flow format, thereby resulting in a short mixing time by diffusion. A good linear relationship was obtained between the resorufin concentration up to 30 mu M and fluorescent intensity over the chip's circular chamber area. The EROD activity was determined with 3-methylcholanthrene (3-MC)-induced hepatocytes. The inhibition effect of a-naphthoflavone was also examined on EROD activity resulting in an IC(50) value of 12.98 mu M.

引用

页码：1282 / 1287

页数：6

共 29 条

[1] Heteroactivation of cytochrome P450 1A1 by teas and tea polyphenols [J].

Anger, DL ;

Petre, MA ;

Crankshaw, DJ .

BRITISH JOURNAL OF PHARMACOLOGY, 2005, 145 (07) :926-933

[2] Xenobiotic metabolism by cultured primary porcine hepatocytes [J].

Behnia, K ;

Bhatia, S ;

Jastromb, N ;

Balis, U ;

Sullivan, S ;

Yarmush, M ;

Toner, M .

TISSUE ENGINEERING, 2000, 6 (05) :467-479

[3] Microassay for rapid measurement of 7-ethoxyresorufin-O-deethylase activity in intact fish hepatocytes [J].

Behrens, A ;

Schirmer, K ;

Bols, NC ;

Segner, H .

MARINE ENVIRONMENTAL RESEARCH, 1998, 46 (1-5) :369-373

[4]

BURKE MD, 1974, DRUG METAB DISPOS, V2, P583

[5] Evaluation of a microfluidic based cell culture platform with primary human hepatocytes for the prediction of hepatic clearance in human [J].

Chao, P. ;

Maguire, T. ;

Novik, E. ;

Cheng, K. -C. ;

Yarmush, M. L. .

BIOCHEMICAL PHARMACOLOGY, 2009, 78 (06) :625-632

[6] Single-point sampling for assessment of constitutive, induced, and inhibited cytochrome P450 3A activity with alfentanil or midazolam [J].

Chaobal, HN ;

Kharasch, ED .

CLINICAL PHARMACOLOGY & THERAPEUTICS, 2005, 78 (05) :529-539

[7]

CHARON L, 2008, ENCY MICRONANOFLUIDI, V3, P1851

[8] INHIBITION OF MONOOXYGENASE ACTIVITIES IN HUMAN HEPATOCYTES BY INTERFERONS [J].

DONATO, MT ;

HERRERO, E ;

GOMEZLECHON, MJ ;

CASTELL, JV .

TOXICOLOGY IN VITRO, 1993, 7 (04) :481-485

[9]

Fick A., 1855, ANN PHYS, V170, P59, DOI [DOI 10.1002/ANDP.18551700105, 10.1002/andp.18551700105]

[10] Hepatocytes -: the choice to investigate drug metabolism and toxicity in man:: In vitro variability as a reflection of in vivo [J].

Gomez-Lechon, Maria Jose ;

Castell, Jose Vicente ;

Donato, Maria Teresa .

CHEMICO-BIOLOGICAL INTERACTIONS, 2007, 168 (01) :30-50

← 1 2 3 →