Work

Protocol for perfusing human axillary lymph nodes ex vivo to study structure and function in real time

Year: 2025

Type: article

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 • Instructions for perfusing human lymph nodes ex vivo to maintain viability for 24 h • Steps to measure real-time flow rates, acid-base balance, and perfusion pressures • Can be used to study lymph node structure and function in different disease states • Potential to test drug responses and discover biomarkers in patient-derived tissue Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103624

Authors: Amy Llewellyn, Rachel Barrow-McGee, Julia Stevenson, Jasmine Gore, Kalnisha Naidoo