Derivation of functional thymic epithelial organoid lines from adult murine thymus
Summary: 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,...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-04-01
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| Series: | Cell Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124724003474 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Sangho Lim |
| collection | DOAJ |
| description | Summary: 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. |
| first_indexed | 2024-04-24T16:50:31Z |
| format | Article |
| id | doaj.art-4e939fc710f34c5baa9c4a569aa45c17 |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-04-24T16:50:31Z |
| publishDate | 2024-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-4e939fc710f34c5baa9c4a569aa45c172024-03-29T05:49:55ZengElsevierCell Reports2211-12472024-04-01434114019Derivation of functional thymic epithelial organoid lines from adult murine thymusSangho Lim0Gijs J. F. van Son1Ni Luh Wisma Eka Yanti2Amanda Andersson-Rolf3Sam Willemsen4Jeroen Korving5Hong-Gyun Lee6Harry Begthel7Hans Clevers8Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, Utrecht 3584 CT, the Netherlands; Oncode Institute, Utrecht, the NetherlandsOncode Institute, Utrecht, the Netherlands; The Princess Máxima Center for Pediatric Oncology, Utrecht 3584 CS, the NetherlandsHubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, Utrecht 3584 CT, the Netherlands; Oncode Institute, Utrecht, the NetherlandsHubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, Utrecht 3584 CT, the Netherlands; Oncode Institute, Utrecht, the NetherlandsHubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, Utrecht 3584 CT, the Netherlands; Oncode Institute, Utrecht, the NetherlandsHubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, Utrecht 3584 CT, the Netherlands; Oncode Institute, Utrecht, the NetherlandsAnn Romney Center for Neurologic Diseases, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USAHubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, Utrecht 3584 CT, the Netherlands; Oncode Institute, Utrecht, the NetherlandsHubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, Utrecht 3584 CT, the Netherlands; Oncode Institute, Utrecht, the Netherlands; The Princess Máxima Center for Pediatric Oncology, Utrecht 3584 CS, the Netherlands; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2211124724003474CP: ImmunologyCP: Stem cell research |
| spellingShingle | 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 Derivation of functional thymic epithelial organoid lines from adult murine thymus Cell Reports CP: Immunology CP: Stem cell research |
| title | Derivation of functional thymic epithelial organoid lines from adult murine thymus |
| title_full | Derivation of functional thymic epithelial organoid lines from adult murine thymus |
| title_fullStr | Derivation of functional thymic epithelial organoid lines from adult murine thymus |
| title_full_unstemmed | Derivation of functional thymic epithelial organoid lines from adult murine thymus |
| title_short | Derivation of functional thymic epithelial organoid lines from adult murine thymus |
| title_sort | derivation of functional thymic epithelial organoid lines from adult murine thymus |
| topic | CP: Immunology CP: Stem cell research |
| url | http://www.sciencedirect.com/science/article/pii/S2211124724003474 |
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