Decellularized lymph node sections with preserved extracellular matrix for stromal cell culture

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

• Level A · Stronger Clinical Evidence
• Level B · Emerging clinical evidence with positive signals
• Level C · Early human research exploring benefits
• Level D · Scientific groundwork from lab and animal studies
• Emerging · Emerging topic under active research

Abstract (original English)

The lymph node (LN) extracellular matrix (ECM) is produced by stromal cells like fibroblastic reticular cells (FRCs) and supports adaptive immunity by guiding immune cell interactions. Disruption of this ECM in cancer and chronic inflammation has been shown to promote disease progression. While interactions between cells and the LN ECM are critical for immunity, they remain difficult to study due to limitations in current models and reliance on animal studies. To address this, LNs could be decellularized to generate cell-free scaffolds that are subsequently reseeded with cells to study how the native LN microenvironment influences cellular behavior. Existing whole-organ decellularization methods preserve ECM features but yield dense scaffolds that restrict uniform cell seeding, limit nutrient diffusion, and hinder imaging analyses. Here, we present a protocol that combines vibratome sectioning (200-μm slices) with detergent decellularization (0.1% SDS and 1% Triton-X) to generate thin LN slices from mouse and human tissues. Decellularized LNs had comparable collagen and GAG concentrations to native tissue, and immunofluorescence staining showed the presence of other ECM proteins. Decellularized sections sustained 21-day FRC culture, enabled FRC-T cell co-culture, and supported high-resolution imaging and flow cytometric analyses, revealing altered gp38 and PDGFRα expression in

Evidence level

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

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