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

doi:10.1016/j.celrep.2024.114019

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

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

6 Citations (Scopus)

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 language	English
Pages (from-to)	114019
Journal	Cell Reports
Volume	43
Issue number	4
DOIs	https://doi.org/10.1016/j.celrep.2024.114019
Publication status	Published - 23 Apr 2024

Keywords

Animals
Organoids/cytology
Thymus Gland/cytology
Epithelial Cells/cytology
Mice
Cell Differentiation
Mice, Nude
T-Lymphocytes/cytology
Mice, Inbred C57BL
Transcription Factors/metabolism
Forkhead Transcription Factors

Access to Document

10.1016/j.celrep.2024.114019

Cite this

@article{c5f088bdc5284a008444a2d78e6cf24e,

title = "Derivation of functional thymic epithelial organoid lines from adult murine thymus",

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.",

keywords = "Animals, Organoids/cytology, Thymus Gland/cytology, Epithelial Cells/cytology, Mice, Cell Differentiation, Mice, Nude, T-Lymphocytes/cytology, Mice, Inbred C57BL, Transcription Factors/metabolism, Forkhead Transcription Factors",

author = "Sangho Lim and {J F van Son}, Gijs and {Wisma Eka Yanti}, {Ni Luh} and Amanda Andersson-Rolf and Sam Willemsen and Jeroen Korving and Hong-Gyun Lee and Harry Begthel and Hans Clevers",

year = "2024",

month = apr,

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doi = "10.1016/j.celrep.2024.114019",

language = "English",

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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

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

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