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Protocol for perfusing human axillary lymph nodes ex vivo to study structure and function in real time
Amy Llewellyn1, Rachel Barrow-McGee2, Julia Stevenson3
1Translational Pathology, Comprehensive Cancer Centre, School of Cancer and Pharmaceutical Sciences, King's College London, London, UK.
Insights
Researchers developed a new method to keep human lymph nodes alive outside the body for 24 hours. This technique allows for detailed study of lymph node structure and function in various diseases and drug responses.
Area of Science:
- Immunology
- Physiology
- Surgical Techniques
Background:
- Lymph nodes are critical for immune responses and fluid homeostasis.
- Studying human lymph nodes ex vivo is challenging due to their short viability.
- Understanding lymph node dynamics in disease requires advanced experimental models.
Purpose of the Study:
- To present a novel protocol for sustaining patient-derived lymph nodes ex vivo.
- To enable extended study of lymph node structure and function.
- To facilitate research into lymph node changes during disease and drug treatment.
Main Methods:
- Utilizing normothermic perfusion to maintain lymph node viability.
- Describing the setup of thermoregulatory and perfusion circuits.
- Detailing the cannulation and perfusion of human lymph nodes.
Main Results:
- Successful maintenance of human lymph nodes ex vivo for up to 24 hours.
- Demonstration of a reproducible protocol for lymph node perfusion.
- Establishment of a platform for investigating lymph node physiology.
Conclusions:
- The described protocol provides a valuable tool for studying human lymph nodes ex vivo.
- This method can advance research in oncology, immunology, and pharmacology.
- The ex vivo perfusion system supports investigations into disease-related lymph node alterations.
Abstract:
Lymph nodes regulate immunity and maintain fluid balance in health and disease. Here, we present a protocol that uses normothermic perfusion to sustain patient-derived lymph nodes ex vivo for up to 24 h to study their structure and function. We describe steps for setting up both thermoregulatory and perfusion circuits, cannulating human lymph nodes, and perfusion. This protocol can be used to study how human lymph nodes change in cancer and other diseases, and/or in response to perturbations, including drugs. For complete details on the use and execution of this protocol, please refer to Barrow-McGee et al.1.
