Generation of multiciliated cells in functional airway epithelia from human induced pluripotent stem cells.
Despite therapeutic advancement, pulmonary disease still remains a major cause of morbidity and mortality around the world. Opportunities to study human lung disease either in vivo or in vitro are currently limited. Using induced pluripotent stem cells (iPSCs), we generated mature multiciliated cells in a functional airway epithelium. Robust multiciliogenesis occurred when notch signaling was inhibited and was confirmed by (i) the assembly of multiple pericentrin-stained centrioles at the apical surface, (ii) expression of transcription factor forkhead box protein J1, and (iii) presence of multiple acetylated tubulin-labeled cilia projections in individual cells. Clara, goblet, and basal cells were all present, confirming the generation of a complete polarized epithelial-cell layer. Additionally, cAMP-activated and cystic fibrosis transmembrane regulator inhibitor 172-sensitive cystic fibrosis transmembrane regulator currents were recorded in isolated epithelial cells. Our report demonstrating the generation of mature multiciliated cells in respiratory epithelium from iPSCs is a significant advance toward modeling a number of human respiratory diseases in vitro.
Laboratory of Genetics, Salk Institute for Biological Studies, La Jolla, CA 92037.
Amy L
AL
Firth
Carl T
CT
Dargitz
Susan J
SJ
Qualls
Tushar
T
Menon
Rebecca
R
Wright
Oded
O
Singer
Fred H
FH
Gage
Ajai
A
Khanna
Inder M
IM
Verma
D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism; D002923:Cilia / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D004707:Endoderm / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008168:Lung / Q000166:cytology*; D051880:Receptors, Notch / Q000378:metabolism; D015398:Signal Transduction
D051880:Receptors, Notch; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
bronchi; definitive endoderm; differentiation