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Isolation of Murine Lymph Node Stromal Cells
Published on: August 19, 2014
Fluid flow regulates stromal cell organization and CCL21 expression in a tissue-engineered lymph node
Alice A Tomei1, Stefanie Siegert, Mirjam R Britschgi
1Institute of Bioengineering, Ecole Polytechnique Fédérale de Lausanne, Lausanne 1015, Switzerland.
Journal of Immunology (Baltimore, Md. : 1950)
|September 8, 2009
Summary
Lymph flow is crucial for T zone fibroblastic reticular cells (TRCs) in lymph nodes. Slow interstitial flow regulates TRC organization and CCL21 secretion, essential for immune cell trafficking.
Area of Science:
- Immunology
- Cell Biology
- Physiology
Background:
- T zone fibroblastic reticular cells (TRCs) in lymph nodes organize immune responses.
- TRCs guide lymphocyte migration and form 3D networks.
- Lymph node conduits transport fluid, and flow increases during inflammation.
Purpose of the Study:
- To investigate the effect of fluid flow on TRC organization and function.
- To understand the role of interstitial flow in the 3D lymph node T zone stromal network.
Main Methods:
- Regenerated 3D lymph node T zone stromal networks using murine TRC clones in scaffolds.
- Applied slow interstitial flow (1-23 microm/min) to the 3D cultures.
- Blocked lymph flow in vivo in mice to assess CCL21 gene expression.
Main Results:
- 3D environment and slow interstitial flow regulate TRC morphology, organization, and CCL21 secretion.
- CCL21 expression was undetectable without flow in vitro.
- In vivo, blocking lymph flow down-regulated CCL21 gene expression within 2 hours.
Conclusions:
- Lymph flow is a homeostatic regulator of TRC activity.
- Increased lymph flow may serve as an early inflammatory cue, enhancing CCL21 expression by TRCs.
- This enhances immune cell trafficking, lymph sampling, and immune response induction.
