Characterization of novel airway submucosal gland cell models for cystic fibrosis studies.
Cultured airway epithelial cells are widely used in cystic fibrosis (CF) research as in vitro models that mimic the in vivo manifestations of the disease and help to define a specific cellular phenotype. Recently, a number of in vitro studies have used an airway adenocarcinoma cell line, Calu-3 that expresses submucosal gland cell features and significant levels of the wild-type CFTR mRNA and protein. We further characterized previously described CF tracheobronchial gland cell lines, CFSMEo- and 6CFSMEo- and determined that these cell lines are compound heterozygotes for the F508del and Q2X mutations, produce vestigial amounts of CFTR mRNA, and do not express detectable CFTR protein. Electrophysiologically, both cell lines are characteristically CF in that they lack cAMP-induced Cl- currents. In this study the cell lines are evaluated in the context of their role as the CF correlate to the Calu-3 cells. Together these cell systems provide defined culture systems to study the biology and pathology of CF. These airway epithelial cell lines may also be a useful negative protein control for numerous studies involving gene therapy by cDNA complementation or gene targeting.
da Paula
Ana Carina
AC
Centre of Human Genetics, National Institute of Health Dr. Ricardo Jorge, Lisboa, Portugal.
Ramalho
Anabela S
AS
Farinha
Carlos M
CM
Cheung
Judy
J
Maurisse
Rosalie
R
Gruenert
Dieter C
DC
Ousingsawat
Jiraporn
J
Kunzelmann
Karl
K
Amaral
Margarida D
MD
D001483:Base Sequence; D002460:Cell Line; D003550:Cystic Fibrosis / Q000473:pathology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D017931:DNA Primers; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D008954:Models, Biological; D009092:Mucous Membrane; D014132:Trachea / Q000033:anatomy & histology / Q000473:pathology*
C505032:CFTR protein, human; D017931:DNA Primers; D019005:Cystic Fibrosis Transmembrane Conductance Regulator