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Characterisation of H441 cells as an in vitro model of distal lung epithelial barrier
Author(s) -
Ehrhardt Carsten,
Gausterer Julia Clara,
Schwagerus Elena,
Salomon Johanna Jessica
Publication year - 2013
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.27.1_supplement.722.1
Subject(s) - organic cation transport proteins , cell culture , chemistry , in vitro , immunocytochemistry , tight junction , biophysics , western blot , chinese hamster ovary cell , symporter , caco 2 , barrier function , microbiology and biotechnology , transporter , human lung , biochemistry , biology , receptor , endocrinology , genetics , gene
No continuously growing cell line of human distal lung epithelial phenotype and with the ability to form polarised, electrically tight monolayers has been described to date. Here, we investigated, if the human lung adenocarcinoma cell line H441 has potential to overcome this issue. Epithelial barrier properties were studied by immunocytochemistry (ICC) against ZO‐1 and measurement of RT. The expression of MDR1 and organic cation/carnitine transporters (OCT/Ns) was also investigated by ICC and immunoblot. Uptake of [ 14 C]‐TEA and [ 3 H]‐acetylcarnitine was carried out to assess OCT/N function. Furthermore, the effects of OCT/N modulators on organic cation uptake were studied. H441 cells formed confluent, electrically tight monolayers with peak RT values of 600–1000 Ω·cm 2 , after 8 days in culture. Presence of P‐gp, OCT1–3, OCTN1 and 2 was confirmed by Western blot and ICC. Uptake of organic cations was time and temperature‐dependent. In the case of [ 14 C]‐TEA, verapamil and MPP + most efficiently inhibited uptake, whilst amantadine and HC‐3 did not show an effect. D‐carnitine and ergothioneine significantly attenuated [ 3 H]‐acetylcarnitine uptake into H441 cells. H441 cells have the ability to form monolayers with appreciable barrier properties. Moreover, their drug transporter expression and activity is consistent with what has been reported for human alveolar epithelial cells in primary culture.