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Organotypic cell culture models of human nasal mucosa for in-vitro drug absorption studies
Project
Project code: BfR-ZEBET-08-1328-469
Contract period: 01.09.2011
- 31.08.2013
Purpose of research: Basic research
The purpose of this project is further characterization of reconstructed organotypic human cell culture model of nasal mucosa as replacement for animal experiments in drug absorption studies in terms of the expression of active transport proteins.
Filter inserts with polyethylen terephthalate seems to be the best filter material for culturing RPMI 2650 cells. Various efflux and uptake assays suggest functional expression of the ABC transporter proteins P-gp and MRP1 in RPMI 2650 cells and human nasal mucosa. These findings could already be verified for P-gp by indirect immunofluorescence staining. Both, RPMI 2650 cells as well as specimens of human nasal mucosa, exhibited PEPT2 function determined by glycyl sarcosine uptake studies. In addition, expression of PEPT2 could be shown in all samples by indirect immunofluorescence staining.The absorbed amounts of trans-4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (Di-ASP) were higher for different cultured RPMI 2650 cells in absence of the inhibitor verapamil. Enhanced uptake of this cationic fluorescent probe suggests activity of organic cation transporters from the SLC subfamily 22. An efflux assay using radiolabeled digoxin and verapamil as unspecific inhibitor confirmed functional expression of P-glycoprotein in RPMI 2650 models as well as specimens of human nasal mucosa. These findings could be verified by indirect immunofluorescence staining for all samples.
Section overview
Subjects
- Biotechnology
Framework programme
Funding programme
Excutive institution
BfR - Centre for Documentation and Evaluation of Alternatives to Animal Experiments (BfR - ZEBET)
