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Updated: Jun 11, 2025
Isolation of Murine Lymph Node Stromal Cells
Published on: August 19, 2014
On-chip human lymph node stromal network for evaluating dendritic cell and T-cell trafficking
Brian J Kwee1,2, Mona Mansouri1, Adovi Akue3
1Division of Cell Therapy 1, Office of Therapeutic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, United States of America.
Insights
Researchers engineered a 3D lymph node stromal network using fibroblastic reticular cells (FRCs). This model demonstrates that FRCs can recruit T-cells and dendritic cells (DCs) independently of CCR7 ligands, offering new insights into immune cell migration.
Area of Science:
- Immunology
- Biomedical Engineering
- Cell Biology
Background:
- The lymph node paracortex (T-cell zone) relies on fibroblastic reticular cells (FRCs) to guide T-cell and dendritic cell (DC) trafficking via chemokine secretion.
- Understanding these migration mechanisms is crucial for immune response modulation and therapeutic development.
Purpose of the Study:
- To engineer a functional 3D lymph node stromal network model using human cultured FRCs.
- To investigate T-cell and DC migration towards this engineered network, particularly exploring CCR7-independent pathways.
Main Methods:
- Utilized multi-channel microfluidic devices to create a 3D hydrogel matrix (collagen I-fibrin) embedding human FRCs.
- Characterized the FRC network's self-assembly, extracellular matrix secretion, and chemokine expression.
- Assessed the migration of monocyte-derived DCs and T-cells (CD45RA+, CD45RA-) towards the engineered network compared to a blank hydrogel.
Main Results:
- Engineered FRCs self-assembled into an interconnected network, secreting key extracellular matrix proteins and various chemokines.
- Despite lacking characteristic CCR7-ligand chemokines (CCL19, CCL21), the FRC network significantly enhanced migration of DCs and T-cells.
- The FRC network demonstrated co-recruitment of DCs and antigen-specific T-cells, indicating functional immune cell interactions.
Conclusions:
- The engineered 3D lymph node stromal network effectively models key aspects of the paracortex microenvironment.
- This model highlights the capacity of FRCs to recruit immune cells via CCR7-independent mechanisms.
- The platform offers a valuable tool for studying human DC and T-cell migration dynamics in lymph nodes.
Abstract:
The lymph node paracortex, also known as the T-cell zone, consists of a network of fibroblastic reticular cells (FRCs) that secrete chemokines to induce T-cell and dendritic cell (DC) trafficking into the paracortex. To model the lymph node paracortex, we utilize multi-channel microfluidic devices to engineer a 3D lymph node stromal network from human cultured FRCs embedded in a collagen I-fibrin hydrogel. In the hydrogel, the FRCs self-assemble into an interconnected network, secrete the extracellular matrix proteins entactin, collagen IV, and fibronectin, as well as express an array of immune cell trafficking chemokines. Although the engineered FRC network did not secrete characteristic CCR7-ligand chemokines (i.e. CCL19 and CCL21), human primary TNF-αmatured monocyte-derived DCs, CD45RA+T-cells, and CD45RA-T-cells migrate toward the lymph node stromal network to a greater extent than toward a blank hydrogel. Furthermore, the FRCs co-recruit DCs and antigen-specific T-cells into the lymph node stromal network. This engineered lymph node stromal network may help evaluate how human DCs and T-cells migrate into the lymph node paracortex via CCR7-independent mechanisms.
