title: Efficient generation of functional CFTR-expressing airway epithelial cells from human pluripotent stem cells.
abstract: Airway epithelial cells are of great interest for research on lung development, regeneration and disease modeling. This protocol describes how to generate cystic fibrosis (CF) transmembrane conductance regulator protein (CFTR)-expressing airway epithelial cells from human pluripotent stem cells (PSCs). The stepwise approach from PSC culture to differentiation into progenitors and then mature epithelia with apical CFTR activity is outlined. Human PSCs that were inefficient at endoderm differentiation using our previous lung differentiation protocol were able to generate substantial lung progenitor cell populations. Augmented CFTR activity can be observed in all cultures as early as at 35 d of differentiation, and full maturation of the cells in air-liquid interface cultures occurs in <5 weeks. This protocol can be used for drug discovery, tissue regeneration or disease modeling.
authors: Program in Developmental and Stem Cell Biology, Hospital for Sick Children, Toronto, Ontario, Canada.
authors: Amy P
authors: AP
authors: Wong
authors: Program in Molecular Structure and Function, Hospital for Sick Children, Toronto, Ontario, Canada.
authors: Stephanie
authors: S
authors: Chin
authors: Program in Molecular Structure and Function, Hospital for Sick Children, Toronto, Ontario, Canada.
authors: Sunny
authors: S
authors: Xia
authors: Program in Developmental and Stem Cell Biology, Hospital for Sick Children, Toronto, Ontario, Canada.
authors: Jodi
authors: J
authors: Garner
authors: Program in Molecular Structure and Function, Hospital for Sick Children, Toronto, Ontario, Canada.
authors: Christine E
authors: CE
authors: Bear
authors: 1] Program in Developmental and Stem Cell Biology, Hospital for Sick Children, Toronto, Ontario, Canada. [2] Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada. [3] Department of Obstetrics and Gynaecology, University of Toronto, Toronto, Ontario, Canada.
authors: Janet
authors: J
authors: Rossant
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism*; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology*; D020545:Respiratory Mucosa / Q000166:cytology* / Q000378:metabolism
chemical_list: C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
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