FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors

Chew-Li Soh, Antonietta Giudice, Robert Alexander Jenny, David A Elliott, Tanya Hatzistavrou, Suzanne J Micallef, Korosh Kianizad, Natalie L Seach, Juan Carlos Zuniga-Pflucker, Ann P Chidgey, Alan O Trounson, Susie [Susan] K Nilsson, David Norman Haylock, Richard L Boyd, Andrew G Elefanty, Edouard G Stanley

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16 Citations (Scopus)

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-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.
Original languageEnglish
Pages (from-to)925 - 937
Number of pages13
JournalStem Cell Reports
Volume2
Issue number6
DOIs
Publication statusPublished - 2014

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