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

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 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.