Characterization of immortal cystic fibrosis tracheobronchial gland epithelial cells.

Abstract

Tracheobronchial glands were isolated and cultured from a patient with cystic fibrosis (CF). Cultured epithelial cells were transformed with pSVori-. All transformed cell lines express cytokeratin filaments and at early passages express the junctional complex molecule cell CAM 120/80, indicating their epithelial origin. Several gland cell lines express antigens that localize to secretory cells in vivo. Cl- transport measured by 36Cl efflux shows that CF gland epithelial cells, like CF surface airway and nasal polyp epithelial cells, are unable to respond to increases in intracellular cAMP. However, they do produce an increase in intracellular cAMP after treatment with isoproterenol or forskolin. One CF gland cell line shows increased intracellular calcium in response to a number of agents and increased Cl- efflux comparable to that observed in a non-CF airway surface epithelial cell line after addition of calcium ionophore. All cell lines express CF transmembrane conductance regulator mRNA, as measured by PCR amplification of first-strand cDNA. The CF tracheobronchial gland cell lines described here are compound heterozygotes, having a single copy of the delta F508 mutation.

Caspases shutdown nonsense-mediated mRNA decay during apoptosis

“…Immortalized cystic fibrosis airway epithelial cells, IB3 harboring the W1282X nonsense mutation in CFTR gene or, alternatively, 6CFSMEo-harboring the Q2X nonsense mutation in CFTR gene (Supplementary Figure S3) 27,28 were also assayed. We first determined that 2 μM staurosporine was the lowest concentration that provided efficient apoptosis in each cell line (Supplementary Figure S3A) even though this level of apoptosis is about two-fold less than that obtained in HeLa cells.…”

Poly (D, L-lactide-co-glycolide)/DNA microspheres to facilitate prolonged transgene expression in airway epithelium in vitro, ex vivo and in vivo

“…The SV-transformed epithelial cell line Cos 7 31 was maintained at 371C/5% CO 2 in Dulbecco's modified Eagles medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% L-glutamine and 1% penicillin/ streptomycin. The CF submucosal epithelial cell line 32 were maintained in minimal essential medium (MEM) supplemented with 10% FBS, 1% penicillin/streptomycin, 1% L-glutamine and nonessential amino acids.…”

The effect of mucolytic agents on gene transfer across a CF sputum barrier in vitro

“…This study, however, bovine serum (FBS), 1% l-glutamine and 1% penicillin/streptomycin. The human bronchial epithelial demonstrates a beneficial effect of rhDNase upon gene transfer efficiency across a sputum barrier, albeit only cell line (16HBE14o−) 32 and the CF submucosal epithelial cell line (2CFSMEo−) 33 were maintained in minimum with partial restoration. The observations suggest, furthermore, that neutrophil-derived DNA may bind to epiessential medium (MEM) supplemented with 10% FBS, 1% penicillin/streptomycin, 1% l-glutamine and nonthelial cell surfaces in a manner that is not reversible by PBS washing and that it is this sputum component which essential amino acids.…”