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

Area of Science:

• Immunology
• Biomaterials Science
• Cell Biology

Background:

• The lymph node (LN) extracellular matrix (ECM) is crucial for adaptive immunity, guiding immune cell interactions.
• ECM disruption in cancer and inflammation promotes disease progression.
• Studying LN ECM-cellular interactions is challenging due to model limitations and reliance on animal studies.

Purpose of the Study:

• To develop a novel method for creating thin, cell-free lymph node scaffolds for studying the native microenvironment.
• To overcome limitations of existing whole-organ decellularization techniques that hinder cell seeding, nutrient diffusion, and imaging.

Main Methods:

• Combined vibratome sectioning (200-μm slices) with detergent decellularization (0.1% SDS, 1% Triton-X) on mouse and human LNs.
• Characterized decellularized scaffolds for ECM protein content (collagen, GAGs, other ECM proteins).
• Cultured fibroblastic reticular cells (FRCs) on scaffolds for 21 days and performed FRC-T cell co-cultures.

Main Results:

• Decellularized LN slices retained comparable collagen and glycosaminoglycan (GAG) concentrations to native tissue.
• Immunofluorescence confirmed the presence of various ECM proteins in the decellularized scaffolds.
• Scaffolds supported long-term FRC culture, FRC-T cell co-culture, and high-resolution imaging.
• Revealed altered gp38 and PDGFRα expression in FRCs cultured on scaffolds compared to 2D cultures.

Conclusions:

• Thin, decellularized LN slices provide a viable model for studying LN ECM-cellular interactions.
• This method facilitates advanced analyses like high-resolution imaging and flow cytometry.
• The developed scaffolds offer a promising platform for investigating immune cell behavior within a biomimetic LN microenvironment.