PURPOSE: To develop a cell culture model of human alveolar epithelial cells in primary culture for the in vitro study of pulmonary absorption and transport. METHODS: Type II pneumocytes isolated from normal human distal lung tissue by enzyme treatment and subsequent purification were plated on fibronectin/collagen coated polyester filter inserts, and cultured using a low-serum growth medium. Characterization of the cell culture was achieved by bioelectric measurements, cell-specific lectin binding, immunohistochemical detection of cell junctions, and by assessment of transepithelial transport of dextrans of varying molecular weights. RESULTS: In culture, the isolated cells spread into confluent monolayers, exhibiting peak transepithelial resistance of 2,180 +/- 62 ohms x cm2 and potential difference of 13.5 +/- 1.0 mV (n = 30-48), and developing tight junctions as well as desmosomes. As assessed by lectin-binding, the cell monolayers consisted of mainly type I cells with some interspersed type II cells, thus well mimicking the situation in vivo. The permeability of hydrophilic macromolecular FITC-dextrans across the cell monolayer was found to be inversely related to their molecular size, with Papp values ranging from 1.7 to 0.2 x 10(-8) cm/sec. CONCLUSIONS: A primary cell culture model of human alveolar epithelial cells has been established, which appears to be a valuable in vitro model for pulmonary drug delivery and transport studies.
PURPOSE: To develop a cell culture model of human alveolar epithelial cells in primary culture for the in vitro study of pulmonary absorption and transport. METHODS: Type II pneumocytes isolated from normal human distal lung tissue by enzyme treatment and subsequent purification were plated on fibronectin/collagen coated polyester filter inserts, and cultured using a low-serum growth medium. Characterization of the cell culture was achieved by bioelectric measurements, cell-specific lectin binding, immunohistochemical detection of cell junctions, and by assessment of transepithelial transport of dextrans of varying molecular weights. RESULTS: In culture, the isolated cells spread into confluent monolayers, exhibiting peak transepithelial resistance of 2,180 +/- 62 ohms x cm2 and potential difference of 13.5 +/- 1.0 mV (n = 30-48), and developing tight junctions as well as desmosomes. As assessed by lectin-binding, the cell monolayers consisted of mainly type I cells with some interspersed type II cells, thus well mimicking the situation in vivo. The permeability of hydrophilic macromolecular FITC-dextrans across the cell monolayer was found to be inversely related to their molecular size, with Papp values ranging from 1.7 to 0.2 x 10(-8) cm/sec. CONCLUSIONS: A primary cell culture model of human alveolar epithelial cells has been established, which appears to be a valuable in vitro model for pulmonary drug delivery and transport studies.
Authors: A Masuda; M Kondo; T Saito; Y Yatabe; T Kobayashi; M Okamoto; M Suyama; T Takahashi; T Takahashi Journal: Cancer Res Date: 1997-11-01 Impact factor: 12.701
Authors: Jennifer Fiegel; Carsten Ehrhardt; Ulrich Friedrich Schaefer; Claus-Michael Lehr; Justin Hanes Journal: Pharm Res Date: 2003-05 Impact factor: 4.200
Authors: Nazanin R Yacobi; Harish C Phuleria; Lucas Demaio; Chi H Liang; Ching-An Peng; Constantinos Sioutas; Zea Borok; Kwang-Jin Kim; Edward D Crandall Journal: Toxicol In Vitro Date: 2007-04-27 Impact factor: 3.500
Authors: Helen J Rippon; Síle Lane; Mingde Qin; N-Siti Ismail; Michael R Wilson; Masao Takata; Anne E Bishop Journal: Proc Am Thorac Soc Date: 2008-08-15
Authors: Marybeth Langer; Elizabeth Stewart Duggan; John Leland Booth; Vineet Indrajit Patel; Ryan A Zander; Robert Silasi-Mansat; Vijay Ramani; Tibor Zoltan Veres; Frauke Prenzler; Katherina Sewald; Daniel M Williams; Kenneth Mark Coggeshall; Shanjana Awasthi; Florea Lupu; Dennis Burian; Jimmy Dale Ballard; Armin Braun; Jordan Patrick Metcalf Journal: Infect Immun Date: 2012-10-01 Impact factor: 3.441
Authors: Jessica Hoppstädter; Britta Diesel; Robert Zarbock; Tanja Breinig; Dominik Monz; Marcus Koch; Andreas Meyerhans; Ludwig Gortner; Claus-Michael Lehr; Hanno Huwer; Alexandra K Kiemer Journal: Respir Res Date: 2010-09-15