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Establishment of a microwell-array-based miniaturized thymic organoid model suitable for high-throughput applications

Abstract:
T cell development depends critically on the thymic stroma-in particular, the diverse array of functionally distinct thymic epithelial cell (TEC) types. However, a robust in vitro thymus model mimicking the native thymus and compatible with medium-/high-throughput analyses is currently lacking. Here, we demonstrate a high-density microwell-array-based miniaturized thymus organoid (mTO) model that supports T cell commitment and development, possesses key organizational characteristics of the native thymus, and is compatible with live imaging and medium-/high-throughput applications. We establish the minimum cellular input required for a functional mTO and show that mTO TEC phenotype and complexity closely mirror those of the native thymus. Finally, we use an mTO to probe the role of fetal thymic mesenchyme, revealing a requirement beyond maintenance of Foxn1 in differentiation/maintenance of mature TEC sub-populations. Collectively, mTOs present an in vitro  model of the native thymus adaptable to medium-/high-throughput applications and validated for exploration of thymus and thymus organoid biology.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1016/j.celrep.2025.115579

Authors

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Institution:
University of Oxford
Division:
MPLS
Department:
Mathematical Institute
Role:
Author
ORCID:
0000-0002-5036-6073


More from this funder
Funder identifier:
https://ror.org/019w4f821
Grant:
602587
Programme:
Seventh Framework Programme (FP7/2007–2013) project ThymiStem
More from this funder
Funder identifier:
https://ror.org/029chgv08
Grant:
211944/Z/18/Z
More from this funder
Funder identifier:
https://ror.org/001aqnf71
Grant:
MR/N013166/1
BB/MO16412/1
NC/X002470/1
MR/T029765/1


Publisher:
Cell Press
Journal:
Cell Reports More from this journal
Volume:
44
Issue:
5
Article number:
115579
Place of publication:
United States
Publication date:
2025-04-15
Acceptance date:
2025-03-27
DOI:
EISSN:
2211-1247
ISSN:
2639-1856
Pmid:
40244847


Language:
English
Keywords:
Pubs id:
2120559
Local pid:
pubs:2120559
Deposit date:
2025-05-16
ARK identifier:

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