Protocol for the generation and differentiation of thymic epithelial organoids from adult mouse thymus tissue

Sam Willemsen, Ni Luh Wisma Eka Yanti, Johan H. van Es, Hans Clevers*, Sangho Lim*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Here, we present a protocol for generating long-term expandable thymic epithelial cell (TEC) organoids from adult mouse thymus. We describe the steps for tissue isolation, dissociation, 3D culture matrix embedding, organoid maintenance, and passaging. We then detail procedures for TEC differentiation into cortical and medullary TEC lineages using defined growth factors, and techniques for analyzing organoids, including flow cytometry and qPCR. This protocol provides a valuable tool for studying TEC development and function. For complete details on the use and execution of this protocol, please refer to Lim et al.1

Original languageEnglish
Article number103883
JournalSTAR protocols
Volume6
Issue number2
DOIs
Publication statusPublished - 20 Jun 2025

Keywords

  • Cell Biology
  • Cell culture
  • Molecular Biology
  • Organoids

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