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

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.