Isolation and air–liquid interface culture of human large airway and bronchiolar epithelial cells


      This article describes the techniques of isolation and culture of human airway epithelial cells from large airways and from distal airways. Both cell types are obtained from lung pieces collected during surgery. The protocols start with an initial step of washing and dissection of the lung pieces to separate large airways or distal airways from the surrounding parenchyma. The second step is enzymatic isolation of epithelial cells from the dissected large or distal airways. Cells are then collected by centrifugation and then seeded onto collagen surfaces. Epithelial cells can be grown at an air–liquid interface and usually form a confluent and functional epithelial layer within days.



        • Blouquit S.
        • Morel H.
        • Hinnrasky J.
        • Naline E.
        • Puchelle E.
        • Chinet T.
        Characterization of ion and fluid transport in human bronchioles.
        Am. J. Respir. Cell Mol. Biol. 2002; 27: 503-510
        • Garcia J.R.
        • Jaumann F.
        • Schulz S.
        • Krause A.
        • Rodriguez-Jimenez J.
        • Forssmann U.
        • et al.
        Identification of a novel, multifunctional beta-defensin (human beta-defensin 3) with specific antimicrobial activity. Its interaction with plasma membranes of Xenopus oocytes and the induction of macrophage chemoattraction.
        Cell Tissue Res. 2001; 306: 257-264
        • Bals R.
        • Xiao W.
        • Sang N.
        • Weiner D.J.
        • Meegalla R.L.
        • Wilson J.M.
        Transduction of well-differentiated airway epithelium by recombinant adeno-associated virus is limited by vector entry.
        J. Virol. 1999; 73: 6085-6088