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Isolation of Murine Lymph Node Stromal Cells
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Isolation of Murine Lymph Node Stromal Cells

Published on: August 19, 2014

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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.

Biofabrication
|September 27, 2024
PubMed
Summary

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.

Keywords:
T-cellsdendritic cellsfibroblastic reticular cellslymph node paracortexmicrophysiological systems

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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.