Applications of Epithelial Cell Culture in Studies of Drug Transport

Tavelin, Staffan; Gråsjö, Johan; Taipalensuu, Jan; Ocklind, Göran; Artursson, Per

doi:10.1385/1-59259-185-x:233

Cited by 83 publications

(141 citation statements)

References 48 publications

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“…The formation of a tight junction is not always in proportion to the expression level of tight junction-associated proteins, and other tight junction-associated proteins such as claudins might contribute to the formation of a tight junction (Furuse et al, 1998;Morita et al, 1999). TEER values were reported to vary according to different clones or passage numbers of Caco-2 cells with a range from 62 to 1290 ⍀ ϫ cm 2 , and the TEER value at 30 to 40 passages, which was close to the passage number of Caco-2 cells in our studies, was approximately 350 ⍀ ϫ cm 2 (Delie and Rubas, 1997;Tavelin et al, 2002). Therefore, the TEER value of 441 ⍀ ϫ cm 2 at 2 weeks in this study was considered to be sufficiently high.…”

Section: Discussionsupporting

confidence: 84%

“…The conventional culture period of Caco-2 cells for transport studies is 21 days, judging from the membrane integrity as assessed by the measurement of transepithelium electrical resistance (TEER) and the permeability of hydrophilic molecules such as mannitol (Artursson and Karlsson, 1991;Tavelin et al, 2002). The culture periods are considered to be important for the formation of a tight junction (Sambuy et al, 2005).…”

mentioning

confidence: 99%

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Culture Period-Dependent Change of Function and Expression of ATP-Binding Cassette Transporters in Caco-2 Cells

Kamiyama¹,

Sugiyama²,

Nakai³

et al. 2009

Drug Metab Dispos

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ABSTRACT:The objective of this study was to determine an appropriate culture period to assess whether a compound of interest is transported by efflux transporters such as human multidrug resistance 1 (hMDR1), human multidrug resistance-associated protein 2 (hMRP2), and human breast cancer resistance protein (hBCRP) in Caco-2 cells. Caco-2 cells were cultured on a Transwell for 1 to 6 weeks. The expression of these transporters in the mRNA and protein levels was examined using a real-time polymerase chain reaction and Western blotting, respectively. Transcellular transport activities using digoxin, ochratoxin A, olmesartan, and estrone-3-sulfate were also examined. Except for digoxin, the permeability coefficient (P app ) ratio of the three compounds at 2 weeks was the highest in the periods tested. The P app ratio of digoxin at 2 weeks was higher than that at 3 weeks. The temporal expression profile of each transporter in the mRNA level was similar to that in the protein level, and the functions of hMRP2 and hBCRP were roughly correlated with the expression in the mRNA and protein levels, but that of hMDR1 was not. These data suggest that among all the culture periods evaluated a 2-week culture is the best culture period for transport studies to identify whether a compound is a substrate for hMDR1, hMRP2, and hBCRP.

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Section: Discussionsupporting

confidence: 84%

mentioning

confidence: 99%

Culture Period-Dependent Change of Function and Expression of ATP-Binding Cassette Transporters in Caco-2 Cells

Kamiyama¹,

Sugiyama²,

Nakai³

et al. 2009

Drug Metab Dispos

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“…The inserts were washed twice with HBSS and incubated in this solution for 15 min at 37˚C. After the placement of microporous inserts into fresh wells, the study of transport started, according to the standard protocol ( 16 ) . The transport solutions were added into the apical compartment (0.5 mL) or the basolateral compartment (1 mL) of the culture chamber.…”

Section: Vitaminsmentioning

confidence: 99%

“…vitamin C ( 15 )). The mechanisms of transport in epithelial cells are recognized as passive transcellular, paracellular, active transcellular (transporter-mediated) or transcytosic ( 16 ). The same substance may be transported passively or actively depending on different factors ( 17 -19 ).…”

mentioning

confidence: 99%

Transport of High Concentration of Thiamin, Riboflavin and Pyridoxine across Intestinal Epithelial Cells Caco-2

Zielińska‐Dawidziak

Grajek

Olejnik

et al. 2008

J Nutr Sci Vitaminol

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SummaryIn this study, we examined the intestinal uptake of thiamin (vitamin B 1 ), riboflavin (vitamin B 2 ) and pyridoxine (vitamin B 6 ) administered at high concentration using intestinal epithelial Caco-2 cells as an in vitro model of drugs and food absorption. The effect of vitamin concentration, culture age, transport direction and incubation temperature on vitamin transport was determined. The vitamin transport was expressed as an apparent permeability coefficient and changes in cumulative fraction transported across epithelial membrane in time. It was found that transepithelial transport of these vitamins is dependent on the experimental factors. At low concentrations an active transport mechanism was observed, whereas at high vitamin concentration a passive transport dominated. At high vitamin concentration the transepithelial flux of vitamins in both directions was similar, which proves the mechanism of passive transport.

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“…sphere with 10% CO 2 ; the medium was changed every second day, and the cells were passaged once a week, as described elsewhere (22). The transport and metabolism studies were performed on Caco-2 cell monolayers grown on cell culture inserts as a model system for the human intestine.…”

mentioning

confidence: 99%

Orally Active Antiviral Tripeptide Glycyl-Prolyl-Glycinamide Is Activated by CD26 (Dipeptidyl Peptidase IV) before Transport across the Intestinal Epithelium

Hubatsch

Lazorova

Vahlne

et al. 2005

Antimicrob Agents Chemother

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The tripeptide amide glycyl-prolyl-glycinamide (GPG-amide) is a new antiretroviral drug candidate, but its absorption mechanism is unknown. In this investigation, the transport and metabolism of GPG-amide were studied in a model of the human intestinal epithelium, Caco-2 cell monolayers. The results show that when the tripeptide amide came into contact with the apical enterocyte membrane, it was degraded by CD26 (dipeptidyl peptidase IV) to glycylproline and the antiretrovirally active metabolite glycinamide. Glycinamide retained antiretroviral activity in vitro after transport through the Caco-2 cell monolayers. The transport of glycinamide across Caco-2 cell monolayers occurred via passive diffusion with an apparent permeability coefficient of about 2 ؋ 10 ؊6 cm s ؊1 , which suggests that it is absorbed by the oral route in sufficient amounts to be considered for oral administration. In conclusion, the tripeptide GPG-amide acts as a prodrug that is activated by CD26 to release the orally active antiretroviral compound glycinamide.

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Applications of Epithelial Cell Culture in Studies of Drug Transport

Cited by 83 publications

References 48 publications

Culture Period-Dependent Change of Function and Expression of ATP-Binding Cassette Transporters in Caco-2 Cells

Culture Period-Dependent Change of Function and Expression of ATP-Binding Cassette Transporters in Caco-2 Cells

Transport of High Concentration of Thiamin, Riboflavin and Pyridoxine across Intestinal Epithelial Cells Caco-2

Orally Active Antiviral Tripeptide Glycyl-Prolyl-Glycinamide Is Activated by CD26 (Dipeptidyl Peptidase IV) before Transport across the Intestinal Epithelium

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