Derivation of functional thymic epithelial organoid lines from adult murine thymus

TY - JOUR

T1 - Derivation of functional thymic epithelial organoid lines from adult murine thymus

AU - Lim, Sangho

AU - J F van Son, Gijs

AU - Wisma Eka Yanti, Ni Luh

AU - Andersson-Rolf, Amanda

AU - Willemsen, Sam

AU - Korving, Jeroen

AU - Lee, Hong-Gyun

AU - Begthel, Harry

AU - Clevers, Hans

N1 - Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.

PY - 2024/4/23

Y1 - 2024/4/23

N2 - 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.

AB - 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.

KW - Animals

KW - Organoids/cytology

KW - Thymus Gland/cytology

KW - Epithelial Cells/cytology

KW - Mice

KW - Cell Differentiation

KW - Mice, Nude

KW - T-Lymphocytes/cytology

KW - Mice, Inbred C57BL

KW - Transcription Factors/metabolism

KW - Forkhead Transcription Factors

U2 - 10.1016/j.celrep.2024.114019

DO - 10.1016/j.celrep.2024.114019

M3 - Article

C2 - 38551965

SN - 2211-1247

VL - 43

SP - 114019

JO - Cell Reports

JF - Cell Reports

IS - 4

ER -