title: FOXN1 (GFP/w) reporter hESCs enable identification of integrin-β4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1(+) thymic epithelial progenitors.
abstract: Thymic epithelial cells (TECs) play a critical role in T cell maturation and tolerance induction. The generation of TECs from in vitro differentiation of human pluripotent stem cells (PSCs) provides a platform on which to study the mechanisms of this interaction and has implications for immune reconstitution. To facilitate analysis of PSC-derived TECs, we generated hESC reporter lines in which sequences encoding GFP were targeted to FOXN1, a gene required for TEC development. Using this FOXN1 (GFP/w) line as a readout, we developed a reproducible protocol for generating FOXN1-GFP(+) thymic endoderm cells. Transcriptional profiling and flow cytometry identified integrin-β4 (ITGB4, CD104) and HLA-DR as markers that could be used in combination with EpCAM to selectively purify FOXN1(+) TEC progenitors from differentiating cultures of unmanipulated PSCs. Human FOXN1(+) TEC progenitors generated from PSCs facilitate the study of thymus biology and are a valuable resource for future applications in regenerative medicine.
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Chew-Li
authors.initials: CL
authors.lastname: Soh
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Antonietta
authors.initials: A
authors.lastname: Giudice
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Jenny
authors.affiliation: Murdoch Childrens Research Institute, The Royal Children's Hospital, Parkville, VIC 3052, Australia.
authors.forename: David A
authors.initials: DA
authors.lastname: Elliott
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Tanya
authors.initials: T
authors.lastname: Hatzistavrou
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Suzanne J
authors.initials: SJ
authors.lastname: Micallef
authors.affiliation: Sunnybrook Research Institute, Department of Immunology, University of Toronto, Toronto, ON M4N 3M5, Canada.
authors.forename: Korosh
authors.initials: K
authors.lastname: Kianizad
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Natalie
authors.initials: N
authors.lastname: Seach
authors.affiliation: Sunnybrook Research Institute, Department of Immunology, University of Toronto, Toronto, ON M4N 3M5, Canada.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Zúñiga-Pflücker
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Ann P
authors.initials: AP
authors.lastname: Chidgey
authors.affiliation: California Institute for Regenerative Medicine, San Francisco, CA 94107, USA.
authors.forename: Alan
authors.initials: A
authors.lastname: Trounson
authors.affiliation: Materials Science and Engineering, Commonwealth Scientific and Industrial Research Organisation, Clayton, VIC 3168, Australia ; Australian Regenerative Medicine Institute, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Susan K
authors.initials: SK
authors.lastname: Nilsson
authors.affiliation: Materials Science and Engineering, Commonwealth Scientific and Industrial Research Organisation, Clayton, VIC 3168, Australia ; Australian Regenerative Medicine Institute, Monash University, Clayton, VIC 3800, Australia.
authors.forename: David N
authors.initials: DN
authors.lastname: Haylock
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Richard L
authors.initials: RL
authors.lastname: Boyd
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia ; Murdoch Childrens Research Institute, The Royal Children's Hospital, Parkville, VIC 3052, Australia.
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Elefanty
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia ; Murdoch Childrens Research Institute, The Royal Children's Hospital, Parkville, VIC 3052, Australia ; Department of Paediatrics, University of Melbourne, Parkville, VIC 3050, Australia.
authors.forename: Edouard G
authors.initials: EG
authors.lastname: Stanley
mesh_terms: D000951:Antigens, Neoplasm / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D000071858:Epithelial Cell Adhesion Molecule; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D051858:Forkhead Transcription Factors / Q000378:metabolism*; D006684:HLA-DR Antigens / Q000378:metabolism*; D006801:Humans; D039663:Integrin beta4 / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology*
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D051858:Forkhead Transcription Factors; D006684:HLA-DR Antigens; D039663:Integrin beta4; C090140:Whn protein