FOXN1GFP/w reporter hESCs enable identification of Integrin-β4, HLA-DR, and EpCAM as markers of human PSC-Derived FOXN1+ thymic epithelial progenitors Academic Article uri icon

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

  • Soh, Chew-Li
  • Giudice, Antonietta
  • Jenny, Robert A
  • Elliott, David A
  • Hatzistavrou, Tanya
  • Micallef, Suzanne J
  • Kianizad, Korosh
  • Seach, Natalie
  • Zúñiga-Pflücker, Juan Carlos
  • Chidgey, Ann P
  • Trounson, Alan
  • Nilsson, Susan K
  • Haylock, David N
  • Boyd, Richard L
  • Elefanty, Andrew G
  • Stanley, Edouard G

publication date

  • 2014