CD90 Marks a Mesenchymal Program in Human Thymic Epithelial Cells In Vitro and In Vivo
Thymic epithelium is critical for the structural integrity of the thymus and for T cell development. Within the fully formed thymus, large numbers of hematopoietic cells shape the thymic epithelium into a scaffold-like structure which bears little similarity to classical epithelial layers, such as t...
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Frontiers Media S.A.
2022-03-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.846281/full |
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| author | Shicheng Sun Shicheng Sun Shicheng Sun Jacky Y. Li Jacky Y. Li Jacky Y. Li Hieu T. Nim Hieu T. Nim Hieu T. Nim Hieu T. Nim Adam Piers Adam Piers Mirana Ramialison Mirana Ramialison Mirana Ramialison Mirana Ramialison Enzo R. Porrello Enzo R. Porrello Enzo R. Porrello Enzo R. Porrello Igor E. Konstantinov Igor E. Konstantinov Igor E. Konstantinov Igor E. Konstantinov Andrew G. Elefanty Andrew G. Elefanty Andrew G. Elefanty Andrew G. Elefanty Edouard G. Stanley Edouard G. Stanley Edouard G. Stanley Edouard G. Stanley |
| author_facet | Shicheng Sun Shicheng Sun Shicheng Sun Jacky Y. Li Jacky Y. Li Jacky Y. Li Hieu T. Nim Hieu T. Nim Hieu T. Nim Hieu T. Nim Adam Piers Adam Piers Mirana Ramialison Mirana Ramialison Mirana Ramialison Mirana Ramialison Enzo R. Porrello Enzo R. Porrello Enzo R. Porrello Enzo R. Porrello Igor E. Konstantinov Igor E. Konstantinov Igor E. Konstantinov Igor E. Konstantinov Andrew G. Elefanty Andrew G. Elefanty Andrew G. Elefanty Andrew G. Elefanty Edouard G. Stanley Edouard G. Stanley Edouard G. Stanley Edouard G. Stanley |
| author_sort | Shicheng Sun |
| collection | DOAJ |
| description | Thymic epithelium is critical for the structural integrity of the thymus and for T cell development. Within the fully formed thymus, large numbers of hematopoietic cells shape the thymic epithelium into a scaffold-like structure which bears little similarity to classical epithelial layers, such as those observed in the skin, intestine or pancreas. Here, we show that human thymic epithelial cells (TECs) possess an epithelial identity that also incorporates the expression of mesenchymal cell associated genes, whose expression levels vary between medullary and cortical TECs (m/cTECs). Using pluripotent stem cell (PSC) differentiation systems, we identified a unique population of cells that co-expressed the master TEC transcription factor FOXN1, as well as the epithelial associated marker EPCAM and the mesenchymal associated gene CD90. Using the same serum free culture conditions, we also observed co-expression of EPCAM and CD90 on cultured TECs derived from neonatal human thymus in vitro. Single cell RNA-sequencing revealed these cultured TECs possessed an immature mTEC phenotype and expressed epithelial and mesenchymal associated genes, such as EPCAM, CLDN4, CD90 and COL1A1. Importantly, flow cytometry and single cell RNA-sequencing analysis further confirmed the presence of an EPCAM+CD90+ population in the CD45- fraction of neonatal human thymic stromal cells in vivo. Using the human thymus cell atlas, we found that cTECs displayed more pronounced mesenchymal characteristics than mTECs during embryonic development. Collectively, these results suggest human TECs possess a hybrid gene expression program comprising both epithelial and mesenchymal elements, and provide a basis for the further exploration of thymus development from primary tissues and from the in vitro differentiation of PSCs. |
| first_indexed | 2024-12-13T15:11:00Z |
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| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-13T15:11:00Z |
| publishDate | 2022-03-01 |
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| series | Frontiers in Immunology |
| spelling | doaj.art-78af4cce953b45bf916d16eb982f3b102022-12-21T23:40:53ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-03-011310.3389/fimmu.2022.846281846281CD90 Marks a Mesenchymal Program in Human Thymic Epithelial Cells In Vitro and In VivoShicheng Sun0Shicheng Sun1Shicheng Sun2Jacky Y. Li3Jacky Y. Li4Jacky Y. Li5Hieu T. Nim6Hieu T. Nim7Hieu T. Nim8Hieu T. Nim9Adam Piers10Adam Piers11Mirana Ramialison12Mirana Ramialison13Mirana Ramialison14Mirana Ramialison15Enzo R. Porrello16Enzo R. Porrello17Enzo R. Porrello18Enzo R. Porrello19Igor E. Konstantinov20Igor E. Konstantinov21Igor E. Konstantinov22Igor E. Konstantinov23Andrew G. Elefanty24Andrew G. Elefanty25Andrew G. Elefanty26Andrew G. Elefanty27Edouard G. Stanley28Edouard G. Stanley29Edouard G. Stanley30Edouard G. Stanley31Murdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaFaculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, AustraliaThe Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Murdoch Children’s Research Institute, Parkville, VIC, AustraliaMurdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaFaculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, AustraliaThe Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Murdoch Children’s Research Institute, Parkville, VIC, AustraliaMurdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaFaculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, AustraliaThe Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Murdoch Children’s Research Institute, Parkville, VIC, AustraliaAustralian Regenerative Medicine Institute and Systems Biology Institute Australia, Monash University, Clayton, VIC, AustraliaMurdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaMelbourne Centre for Cardiovascular Genomics and Regenerative Medicine, Royal Children’s Hospital, Melbourne, VIC, AustraliaMurdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaFaculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, AustraliaThe Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Murdoch Children’s Research Institute, Parkville, VIC, AustraliaAustralian Regenerative Medicine Institute and Systems Biology Institute Australia, Monash University, Clayton, VIC, AustraliaMurdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaFaculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, AustraliaThe Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Murdoch Children’s Research Institute, Parkville, VIC, AustraliaMelbourne Centre for Cardiovascular Genomics and Regenerative Medicine, Royal Children’s Hospital, Melbourne, VIC, AustraliaMurdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaFaculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, AustraliaAustralian Regenerative Medicine Institute and Systems Biology Institute Australia, Monash University, Clayton, VIC, AustraliaDepartment of Cardiac Surgery, Royal Children’s Hospital, Melbourne, VIC, AustraliaMurdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaFaculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, AustraliaThe Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Murdoch Children’s Research Institute, Parkville, VIC, AustraliaDepartment of Anatomy and Developmental Biology, Monash University, Clayton, VIC, AustraliaMurdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, VIC, AustraliaFaculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, AustraliaThe Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Murdoch Children’s Research Institute, Parkville, VIC, AustraliaDepartment of Anatomy and Developmental Biology, Monash University, Clayton, VIC, AustraliaThymic epithelium is critical for the structural integrity of the thymus and for T cell development. Within the fully formed thymus, large numbers of hematopoietic cells shape the thymic epithelium into a scaffold-like structure which bears little similarity to classical epithelial layers, such as those observed in the skin, intestine or pancreas. Here, we show that human thymic epithelial cells (TECs) possess an epithelial identity that also incorporates the expression of mesenchymal cell associated genes, whose expression levels vary between medullary and cortical TECs (m/cTECs). Using pluripotent stem cell (PSC) differentiation systems, we identified a unique population of cells that co-expressed the master TEC transcription factor FOXN1, as well as the epithelial associated marker EPCAM and the mesenchymal associated gene CD90. Using the same serum free culture conditions, we also observed co-expression of EPCAM and CD90 on cultured TECs derived from neonatal human thymus in vitro. Single cell RNA-sequencing revealed these cultured TECs possessed an immature mTEC phenotype and expressed epithelial and mesenchymal associated genes, such as EPCAM, CLDN4, CD90 and COL1A1. Importantly, flow cytometry and single cell RNA-sequencing analysis further confirmed the presence of an EPCAM+CD90+ population in the CD45- fraction of neonatal human thymic stromal cells in vivo. Using the human thymus cell atlas, we found that cTECs displayed more pronounced mesenchymal characteristics than mTECs during embryonic development. Collectively, these results suggest human TECs possess a hybrid gene expression program comprising both epithelial and mesenchymal elements, and provide a basis for the further exploration of thymus development from primary tissues and from the in vitro differentiation of PSCs.https://www.frontiersin.org/articles/10.3389/fimmu.2022.846281/fullhuman thymic epithelial cellsepithelial and mesenchymal componentsprimary cells culturepluripotent stem cell differentiationCD90/Thy1cell identity |
| spellingShingle | Shicheng Sun Shicheng Sun Shicheng Sun Jacky Y. Li Jacky Y. Li Jacky Y. Li Hieu T. Nim Hieu T. Nim Hieu T. Nim Hieu T. Nim Adam Piers Adam Piers Mirana Ramialison Mirana Ramialison Mirana Ramialison Mirana Ramialison Enzo R. Porrello Enzo R. Porrello Enzo R. Porrello Enzo R. Porrello Igor E. Konstantinov Igor E. Konstantinov Igor E. Konstantinov Igor E. Konstantinov Andrew G. Elefanty Andrew G. Elefanty Andrew G. Elefanty Andrew G. Elefanty Edouard G. Stanley Edouard G. Stanley Edouard G. Stanley Edouard G. Stanley CD90 Marks a Mesenchymal Program in Human Thymic Epithelial Cells In Vitro and In Vivo Frontiers in Immunology human thymic epithelial cells epithelial and mesenchymal components primary cells culture pluripotent stem cell differentiation CD90/Thy1 cell identity |
| title | CD90 Marks a Mesenchymal Program in Human Thymic Epithelial Cells In Vitro and In Vivo |
| title_full | CD90 Marks a Mesenchymal Program in Human Thymic Epithelial Cells In Vitro and In Vivo |
| title_fullStr | CD90 Marks a Mesenchymal Program in Human Thymic Epithelial Cells In Vitro and In Vivo |
| title_full_unstemmed | CD90 Marks a Mesenchymal Program in Human Thymic Epithelial Cells In Vitro and In Vivo |
| title_short | CD90 Marks a Mesenchymal Program in Human Thymic Epithelial Cells In Vitro and In Vivo |
| title_sort | cd90 marks a mesenchymal program in human thymic epithelial cells in vitro and in vivo |
| topic | human thymic epithelial cells epithelial and mesenchymal components primary cells culture pluripotent stem cell differentiation CD90/Thy1 cell identity |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.846281/full |
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