Protocol for generating human tonsil immune organoids with epithelial and germinal center layers using low-Matrigel culture

  • STAR Protoc. 2026 Mar 20;7(1):104381. doi: 10.1016/j.xpro.2026.104381.
Dandan Meng  1 ,  Han Ding  1 ,  Qiyu Deng  1 ,  Xuqi Li  1 ,  Yuan Tao  2 ,  Xiao Liu  3
Affiliations
  • 1. Institute of Biopharmaceutical and Health Engineering (iBHE), Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China.
  • 2. Department of Otorhinolaryngology, Peking University Shenzhen Hospital, No. 1120 Lianhua Road, Futian District, Shenzhen 518036, China. Electronic address: [email protected].
  • 3. Institute of Biopharmaceutical and Health Engineering (iBHE), Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China. Electronic address: [email protected].
PMID: 41722051 DOI: 10.1016/j.xpro.2026.104381
Abstract

Tonsil immune organoids traditionally include either epithelial layers or immune compartments, failing to recapitulate the integrated physiological microenvironment of human tonsils. Here, we present a protocol to generate tonsil immune organoids that recapitulate both epithelial and germinal center-like structures using a low-concentration Matrigel transwell system. We describe Organoid assembly, immune stimulation, and evaluation of epithelial architecture and germinal center formation. This platform provides a versatile tool for dissecting human immune responses, assessing vaccines, discovering antibodies, and exploring autoimmune mechanisms.

Keywords
Cell Biology; Flow Cytometry; Immunology; Organoids.
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