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Updated: Jan 10, 2026
Isolation of Murine Lymph Node Stromal Cells
Published on: August 19, 2014
Decellularized lymph node sections with preserved extracellular matrix for stromal cell culture
Estefania Esparza1,2, Leonor N Teles3,4, Alisa Fedotova2
1Department of Biomedical Engineering, University of Miami, Coral Gables, FL, 33146, USA.
Insights
Researchers developed thin, decellularized lymph node (LN) slices to study the extracellular matrix (ECM). This new method enables better cell culture and analysis of immune cell interactions within the native LN microenvironment.
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.
Abstract:
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 FRCs relative to 2D culture.
