title: Isolation and culture of airway epithelial cells from chronically infected human lungs.
abstract: We describe procedures for isolating and culturing airway epithelial cells from chronically infected human lungs. Experience in our laboratory demonstrated the need to balance pathogen eradication against antibiotic toxicity to epithelial cells. To provide a logical basis for antibiotic selection and dose, we systematically analyzed the cytotoxicity of antibiotics useful against typical pathogens. Alone, colistin, ciprofloxacin, doxycycline, and tobramycin were moderately toxic at concentrations close to those used in cell culture, whereas amphotericin, ceftazidime, chloramphenicol, imipenem, meropenem, piperacillin, sulfamethoxazole/trimethoprim, and vancomycin were nontoxic even at concentrations many times the antimicrobial level. Epithelial cytotoxicity of combined antibiotics was additive, with no evidence of competition or synergism. Antibiotics had little effect on initial cell attachment and did not acutely lyse cells, but inhibited subsequent growth. Interestingly, cytotoxicity decreased markedly with increasing epithelial cell density. Cystic fibrosis (CF) and non-CF epithelial cells showed no differences in sensitivity to the antibiotics tested and initial exposure to antibiotics did not affect the electrophysiologic properties of resistance or short circuit current in well-differentiated cells. Tailored combinations of antibiotics at appropriate doses killed even multidrug-resistant bacteria. Thus, epithelial cells can usually be cultured from chronically infected CF airways.
authors.affiliation: Cystic Fibrosis/Pulmonary Research and Treatment Center, The University of North Carolina at Chapel Hill, 27599-7248, USA. randell@med.unc.edu
authors.forename: S H
authors.initials: SH
authors.lastname: Randell
authors.forename: L
authors.initials: L
authors.lastname: Walstad
authors.forename: U E
authors.initials: UE
authors.lastname: Schwab
authors.forename: B R
authors.initials: BR
authors.lastname: Grubb
authors.forename: J R
authors.initials: JR
authors.lastname: Yankaskas
mesh_terms: D000900:Anti-Bacterial Agents / Q000008:administration & dosage* / Q000633:toxicity; D001424:Bacterial Infections / Q000188:drug therapy / Q000473:pathology*; D001980:Bronchi / Q000473:pathology; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002908:Chronic Disease; D003550:Cystic Fibrosis / Q000473:pathology; D018432:Drug Resistance, Multiple; D004359:Drug Therapy, Combination; D004848:Epithelium / Q000473:pathology*; D006801:Humans; D008171:Lung Diseases / Q000382:microbiology / Q000473:pathology*
chemical_list: D000900:Anti-Bacterial Agents