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-beta4 (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.
Soh, C-L., Giudice, A., Jenny, R. A., Elliott, D. A.
, Hatzistavrou, T., Micallef, S. J., ... Stanley, E. G. (2014). FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors
. Stem Cell Reports
(6), 925 - 937. https://doi.org/10.1016/j.stemcr.2014.04.009