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The R3/1 alveolar epithelial type I cell line ‐ Characterisation as an in vitro model for drug disposition
Author(s) -
Ehrhardt Carsten,
Endter Sibylle,
Andersen Rikke,
Koslowski Roland,
Horalkova Lenka
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.762.5
Subject(s) - occludin , in vitro , cell culture , immunofluorescence , tight junction , microbiology and biotechnology , cell , a549 cell , biology , chemistry , biochemistry , antibody , immunology , genetics
The rat cell line, R3/1, has been reported to display several phenotypical features of alveolar epithelial type I cells. In order to characterise the cell line as an in vitro model for pulmonary drug disposition studies, R3/1 cells were cultured on Transwell Clear filters. The transepithelial electrical resistance was measured as a parameter for the integrity of cell layers. Presence of cell junctional proteins (i.e., E‐cadherin, occludin, ZO‐1 and ZO‐2) was studied on mRNA (by reverse transcriptase‐polymerase chain reaction) and antigen levels (by immunofluorescence microscopy), respectively. Furthermore, expression patterns of catabolic peptidases were analysed in R3/1 monolayers. These studies included carboxypeptidase M, aminopeptidases A, B, N and P, γ‐glutamyltransferase, angiotensin‐converting enzyme, dipeptidylpeptidase IV and endopeptidases 24.11 and 24.15. R3/1 cells lack the ability to form confluent monolayers, due to an absence of several structural proteins of the junctional complex (occludin, E‐cadherin). However, the level of catabolic peptidases was found to be similar to that of rat alveolar epithelial type I‐like cells in primary culture. This suggests that R3/1 cells might be a good candidate for in vitro stability assays of protein drugs intended for pulmonary administration.