# Protocol for precision cutting and short-term culture of lymph node tissue slices for modeling of innate immune responses ex vivo

## TL;DR

This paper introduces a protocol for preserving the structure of lymph nodes during short-term culture to study human innate immune responses outside the body.

## Contribution

A novel protocol for precision cutting and ex vivo culture of human lymph node tissue slices to model innate immune responses.

## Key findings

- Lymph node tissue slices can be cultured for up to 20 hours while preserving their microanatomical structure.
- The protocol allows for immune stimuli application and analysis of immune responses using multiple readouts.

## Abstract

Secondary lymphoid organs are highly organized for the initiation of immune responses. Here, we present a protocol that enables this microanatomical structure to be preserved during short-term culture, allowing ex vivo modeling of human innate immune responses. We describe the steps for precision cutting of human lymph nodes into live tissue cross-sections that can be cultured for up to 20 h with immune stimuli and analyzed by multiple readouts.

For complete details on the use and execution of this protocol, please refer to Fergusson et al.1

•Steps for precision cutting of live human lymph node tissue slices•Instructions for short-term culture of tissue slices with innate immune stimuli•Guidance for downstream analyses to determine mechanisms of action of immune stimuli

Steps for precision cutting of live human lymph node tissue slices

Instructions for short-term culture of tissue slices with innate immune stimuli

Guidance for downstream analyses to determine mechanisms of action of immune stimuli

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Secondary lymphoid organs are highly organized for the initiation of immune responses. Here, we present a protocol that enables this microanatomical structure to be preserved during short-term culture, allowing ex vivo modeling of human innate immune responses. We describe the steps for precision cutting of human lymph nodes into live tissue cross-sections that can be cultured for up to 20 h with immune stimuli and analyzed by multiple readouts.