+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

OATP1B1/1B3 activity in plated primary human hepatocytes over time in culture

, : OATP1B1/1B3 activity in plated primary human hepatocytes over time in culture. Biochemical Pharmacology 82(9): 1219-1226

Primary human hepatocytes are widely used as an in vitro model for evaluation of drug metabolism and transport. However, it has been shown that the gene expression of many drug-metabolizing enzymes and transporters change in culture. The aim of the present study was to evaluate the activity of organic anion-transporting polypeptide 1B1 (OATP1B1) and 1B3 (OATP1B3) in plated primary human hepatocytes over time in culture. The uptake kinetics of the OATP1B1/1B3 substrate [(3)H]-estradiol-17β-d-glucuronide was determined in cells from five donors. An extensive and variable decrease in OATP1B1/1B3 activity and/or increase in passive diffusion was observed over time. Already after 6h in culture, the OATP1B1/1B3 activity was not possible to determine in liver cells from one donor, while after 24h, the uptake activity was not measurable in one additional donor. In the other three, the decrease in CL(int) (V(max)/K(m)) values ranged from 15% to 86% after 24h in culture compared to the values measured at 2h. Visual examination of OATP1B1 protein expression by confocal microscopy showed localization to the plasma membrane as expected, and an extensive decrease in OATP1B1 expression over time in culture supported the decline in activity. The significant reduction in SLCO1B1 and SLCO1B3 gene expression over time determined by RT-PCR also supported the loss of OATP1B1/1B3 activity. In conclusion, plated primary human hepatocytes are useful as an in vitro model for OATP1B1/1B3-mediated uptake studies, but the culture time may substantially change the uptake kinetics.


PMID: 21787759

DOI: 10.1016/j.bcp.2011.07.076

Other references

Elbein, A.D.; Mann, R.L.; Renis, H.E.; Stark, W.M.; Koffler, H.; Garner, H.R., 1961: The conversion of D-glucose to 5-keto-6-deoxy-D-arabohexose. Journal of Biological Chemistry 236: 289-292

Fortin, A.; Reid, J.T.; Maiga, A.M.; Sim, D.W.; Wellington, G.H., 1981: Effect of level of energy intake and influence of breed and sex on growth of fat tissue and distribution in the bovine carcass. The rate of deposition of fat, and its distribution among the subcutaneous, intermuscular, internal and intramuscular depots, were studied in 141 cattle, ranging in slaughter weight from 121 to 706 kg. Bulls, steers and heifers of the Holstein-Fre...

Tatsuoka Fumio, 1980: Stress-strain behavior of an idealized anisotropic granular material. Soils and Foundations 20(3): 75-90

Abeku, T.A.; Kristan, M.; Jones, C.; Beard, J.; Mueller, D.H.; Okia, M.; Rapuoda, B.; Greenwood, B.; Cox, J., 2008: Determinants of the accuracy of rapid diagnostic tests in malaria case management: evidence from low and moderate transmission settings in the East African highlands. Background: The accuracy of malaria diagnosis has received renewed interest in recent years due to changes in treatment policies in favour of relatively high-cost artemisinin-based combination therapies. The use of rapid diagnostic tests (RDTs) ba...

Kelso, C.M.; Flammer, P.D.; DeSanto, J.A.; Collins, R.T., 2001: Integral equations applied to wave propagation in two dimensions: modeling the tip of a near-field scanning optical microscope. We present a Green's-function/Green's-theorem integral equation approach to numerically modeling two-dimensional, s-polarized, wave propagation problems effectively for a variety of geometries. The model accurately calculates both near f...

Mckinney T., 1975: Studies on the niche separation in 2 carabid cave beetles. Poulson, Thomas L (Ed ) International Journal Of Speleology, Vol 7, Nos 1-2 Symposium on Life Histories Of Cave Beetles Bloomington, Indiana, U S A , 1973 78p Illus Maps Swets And Zeitlinger B V : Amsterdam, Netherlands Isbn 90-265-0232-X 65-78

Heeg F.J., 1986: Reorganization of electronic hybrid circuit processing method ergonomic layout of a complex work system. Taking as an example the organizational and technical restructuralization of hybrid circuit processing at a medium-sized plant, the various tasks essential in order to solve the problems involved are discussed. The major drawbacks associated with...

Glowacka, D.; Kopacz-Jodczyk, T.; Popowicz, J., 1967: Application of the method of Good and Bessman for estimating glucosamine and galactosamine in tissue hydrolyzates. A purification procedure of amino sugars freed of interfering substances was elaborated. This was achieved by means of Dowex 50X4 (H+) and Dowex 50X8 (NH4+) columns. The purified samples freed of amino acids were desalted on a Dowex 50X4 (H+) colu...

Buttiaux, R.; Zaman, M.A.; Catsaras, M., 1962: Enterobacter in human and animal fecal matter. Annales de L'institut Pasteur 103: 101-103

Zicca A., 1975: Immuno histo cytological methods in the study of thyropathy. Folia Allergologica Et Immunologica Clinica: 338