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ISOLATION AND CULTURE OF AIRWAY EPITHELIAL CELLS FROM CHRONICALLY INFECTED HUMAN LUNGS

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 an... Full description

Journal Title: In vitro cellular & developmental biology. Animal 2001-09, Vol.37 (8), p.480-489
Main Author: RANDELL, SCOTT H
Other Authors: WALSTAD, DIANA L , SCHWAB, UTE E , GRUBB, BARBARA R , YANKASKAS, JAMES R
Format: Electronic Article Electronic Article
Language: English
Subjects:
Publisher: Germany: Society for In Vitro Biology
ID: ISSN: 1071-2690
Link: https://www.ncbi.nlm.nih.gov/pubmed/11669281
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title: ISOLATION AND CULTURE OF AIRWAY EPITHELIAL CELLS FROM CHRONICALLY INFECTED HUMAN LUNGS
format: Article
creator:
  • RANDELL, SCOTT H
  • WALSTAD, DIANA L
  • SCHWAB, UTE E
  • GRUBB, BARBARA R
  • YANKASKAS, JAMES R
subjects:
  • Anti-Bacterial Agents - administration & dosage
  • Anti-Bacterial Agents - toxicity
  • antibiotic resistant bacteria
  • antibiotic toxicity
  • Antibiotics
  • Bacteria
  • Bacterial Infections - drug therapy
  • Bacterial Infections - pathology
  • Bronchi - pathology
  • Cell Adhesion - drug effects
  • CELL AND TISSUE MODELS
  • Cell Count
  • cell culture
  • Cell culture techniques
  • Cell Division - drug effects
  • Cell growth
  • Cell Separation - methods
  • Cell Survival - drug effects
  • Cells, Cultured
  • Chronic Disease
  • Cultured cells
  • Cystic fibrosis
  • Cystic Fibrosis - pathology
  • Cytotoxicity
  • Drug Resistance, Multiple
  • Drug Therapy, Combination
  • Epithelial cells
  • Epithelium - pathology
  • human bronchial epithelial cells
  • Humans
  • Lung Diseases - microbiology
  • Lung Diseases - pathology
  • Lungs
  • Tissue culture techniques
ispartof: In vitro cellular & developmental biology. Animal, 2001-09, Vol.37 (8), p.480-489
description: 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.
language: eng
source:
identifier: ISSN: 1071-2690
fulltext: no_fulltext
issn:
  • 1071-2690
  • 1543-706X
  • 1543-706X
url: Link


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titleISOLATION AND CULTURE OF AIRWAY EPITHELIAL CELLS FROM CHRONICALLY INFECTED HUMAN LUNGS
creatorRANDELL, SCOTT H ; WALSTAD, DIANA L ; SCHWAB, UTE E ; GRUBB, BARBARA R ; YANKASKAS, JAMES R
creatorcontribRANDELL, SCOTT H ; WALSTAD, DIANA L ; SCHWAB, UTE E ; GRUBB, BARBARA R ; YANKASKAS, JAMES R
descriptionWe 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.
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languageeng
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subjectAnti-Bacterial Agents - administration & dosage ; Anti-Bacterial Agents - toxicity ; antibiotic resistant bacteria ; antibiotic toxicity ; Antibiotics ; Bacteria ; Bacterial Infections - drug therapy ; Bacterial Infections - pathology ; Bronchi - pathology ; Cell Adhesion - drug effects ; CELL AND TISSUE MODELS ; Cell Count ; cell culture ; Cell culture techniques ; Cell Division - drug effects ; Cell growth ; Cell Separation - methods ; Cell Survival - drug effects ; Cells, Cultured ; Chronic Disease ; Cultured cells ; Cystic fibrosis ; Cystic Fibrosis - pathology ; Cytotoxicity ; Drug Resistance, Multiple ; Drug Therapy, Combination ; Epithelial cells ; Epithelium - pathology ; human bronchial epithelial cells ; Humans ; Lung Diseases - microbiology ; Lung Diseases - pathology ; Lungs ; Tissue culture techniques
ispartofIn vitro cellular & developmental biology. Animal, 2001-09, Vol.37 (8), p.480-489
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descriptionWe 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.
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0Anti-Bacterial Agents - administration & dosage
1Anti-Bacterial Agents - toxicity
2antibiotic resistant bacteria
3antibiotic toxicity
4Antibiotics
5Bacteria
6Bacterial Infections - drug therapy
7Bacterial Infections - pathology
8Bronchi - pathology
9Cell Adhesion - drug effects
10CELL AND TISSUE MODELS
11Cell Count
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13Cell culture techniques
14Cell Division - drug effects
15Cell growth
16Cell Separation - methods
17Cell Survival - drug effects
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19Chronic Disease
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21Cystic fibrosis
22Cystic Fibrosis - pathology
23Cytotoxicity
24Drug Resistance, Multiple
25Drug Therapy, Combination
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authorRANDELL, SCOTT H ; WALSTAD, DIANA L ; SCHWAB, UTE E ; GRUBB, BARBARA R ; YANKASKAS, JAMES R
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abstractWe 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.
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pubSociety for In Vitro Biology
pmid11669281
doi10.1290/1071-2690(2001)037<0480:IACOAE>2.0.CO;2
tpages10