Derivation of functional thymic epithelial organoid lines from adult murine thymus

Sangho Lim, Gijs J. F. van Son, Ni Luh Wisma Eka Yanti, Amanda Andersson-Rolf, Sam Willemsen, Jeroen Korving, Hong Gyun Lee, Harry Begthel, Hans Clevers*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Thymic epithelial cells (TECs) orchestrate T cell development by imposing positive and negative selection on thymocytes. Current studies on TEC biology are hampered by the absence of long-term ex vivo culture platforms, while the cells driving TEC self-renewal remain to be identified. Here, we generate long-term (>2 years) expandable 3D TEC organoids from the adult mouse thymus. For further analysis, we generated single and double FoxN1-P2A-Clover, Aire-P2A-tdTomato, and Cldn4-P2A-tdTomato reporter lines by CRISPR knockin. Single-cell analyses of expanding clonal organoids reveal cells with bipotent stem/progenitor phenotypes. These clonal organoids can be induced to express Foxn1 and to generate functional cortical- and Aire-expressing medullary-like TECs upon RANK ligand + retinoic acid treatment. TEC organoids support T cell development from immature thymocytes in vitro as well as in vivo upon transplantation into athymic nude mice. This organoid-based platform allows in vitro study of TEC biology and offers a potential strategy for ex vivo T cell development.

Original languageEnglish
Article number114019
JournalCell Reports
Volume43
Issue number4
DOIs
Publication statusPublished - 23 Apr 2024

Keywords

  • Aire
  • CP: Immunology
  • CP: Stem cell research
  • cTEC
  • MHC-II
  • mTEC
  • organoids
  • T cell develpoment
  • thymic epithelial cell
  • thymocytes
  • thymus

Fingerprint

Dive into the research topics of 'Derivation of functional thymic epithelial organoid lines from adult murine thymus'. Together they form a unique fingerprint.

Cite this