Preparation Method and In Vitro Characterization of Nanoparticles Sensitive to Tumor Microenvironment

: Immunotherapy is considered a powerful clinical strategy aiming to boost the immune system to fight cancer. In this context, nanomaterials (NMs) are uniquely suited to improve the development and the broad implementation of cancer immunotherapies by overcoming several challenges. In fact, NMs can be rationally designed to navigate complex physical barriers, respond to tumor microenvironments, and enhance/modulate immune system activation. Here, we present a method to prepare stimuli-responsive biocompatible nanoparticles (NPs) able to target the tumor microenvironment. Moreover, we describe protocols to characterize the physical-chemical properties of NPs as well as to evaluate their biocompatibility and therapeutic potential in vitro on three-dimensional (3D) tumor spheroids.