FOXN1(GFP/w) Reporter hESCs Enable Identification of Integrin-beta 4, HLA-DR, and EpCAM as Markers of Human PSC-Derived FOXNI ( ) Thymic Epithelial Progenitors
Chew-Li Soh, Antonietta Giudice, Robert A Jenny, David A Elliott, Tanya Hatzistavrou, Suzanne J Micallef, Korosh Kianizad, Natalie Seach, Juan Carlos Zuniga-Pfluecker, Ann P Chidgey, Alan Trounson, Susan K Nilsson, David N Haylock, Richard L Boyd, Andrew G Elefanty, Edouard G Stanley
Stem Cell Reports | CELL PRESS | Published : 2014
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 ..View full abstract
The authors thank Robyn Mayberry and the staff of StemCore Victoria for the provision of hESCs and iPSCs and Andrew Fryga and the staff of FlowCore (Monash University) for flow cytometric sorting. They also thank Jade Homann and the staff of Monash University Animal Research Laboratories for assistance with mouse experiments and Dr. Mahmood Mohtashami for technical advice on proT-cell cocultures. This work was supported by grants from the Australian Stem Cell Centre, Stem Cells Australia, the Juvenile Diabetes Research Foundation, the National Health and Medical Research Council of Australia (NHMRC), and the Victoria-California Stem Cell Alliance. K.K. is a recipient of an Alexander Graham Bell Doctoral Award from the Natural Sciences and Engineering Research Council of Canada. J.C.Z.-P. is supported by a Canada Research Chair in Developmental Immunology. S.K.N. and D.N.H. are supported by Australian Research Council Future Fellowships. A.G.E. and E.G.S. are Senior Research Fellows of the NHMRC.