title: Generation of organized anterior foregut epithelia from pluripotent stem cells using small molecules.
abstract: Anterior foregut endoderm (AFE) gives rise to therapeutically relevant cell types in tissues such as the esophagus, salivary glands, lung, thymus, parathyroid and thyroid. Despite its importance, reports describing the generation of AFE from pluripotent stem cells (PSCs) by directed differentiation have mainly focused on the Nkx2.1(+) lung and thyroid lineages. Here, we describe a novel protocol to derive a subdomain of AFE, identified by expression of Pax9, from PSCs using small molecules and defined media conditions. We generated a reporter PSC line for isolation and characterization of Pax9(+) AFE cells, which when transplanted in vivo, can form several distinct complex AFE-derived epithelia, including mucosal glands and stratified squamous epithelium. Finally, we show that the directed differentiation protocol can be used to generate AFE from human PSCs. Thus, this work both broadens the range of PSC-derived AFE tissues and creates a platform enabling the study of AFE disorders.
authors.affiliation: Program in Molecular Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, 368 Plantation Street, Worcester, MA 01605, USA.
authors.forename: Nicola A
authors.initials: NA
authors.lastname: Kearns
authors.forename: Ryan M J
authors.initials: RM
authors.lastname: Genga
authors.forename: Michael
authors.initials: M
authors.lastname: Ziller
authors.forename: Kristina
authors.initials: K
authors.lastname: Kapinas
authors.forename: Heiko
authors.initials: H
authors.lastname: Peters
authors.forename: Michael A
authors.initials: MA
authors.lastname: Brehm
authors.forename: Alexander
authors.initials: A
authors.lastname: Meissner
authors.forename: René
authors.initials: R
authors.lastname: Maehr
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism / Q000637:transplantation; D051542:Hepatocyte Nuclear Factor 3-beta / Q000378:metabolism; D006801:Humans; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism; D051764:PAX9 Transcription Factor / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D054852:Small Molecule Libraries / Q000494:pharmacology*; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors / Q000378:metabolism; D059467:Transcriptome
chemical_list: D003470:Culture Media; C496502:FOXA2 protein, human; C000614414:Nkx2-1 protein, mouse; D009687:Nuclear Proteins; D051764:PAX9 Transcription Factor; C497323:PAX9 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D054852:Small Molecule Libraries; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors; D051542:Hepatocyte Nuclear Factor 3-beta
keywords: AFE; BMPi; DE; ESCs; MEF; PSCs; TGFβi; anterior foregut endoderm; bone morphogenic protein inhibitor; definitive endoderm; embryonic stem cells; mouse embryonic fibroblasts; pluripotent stem cells; transforming growth factor inhibitor