Analyzing the effect of NK cells on in-vivo models of tumor dissemination by digital pathology and flow cytometry.

Despite the significant expansion of the studies aimed at generating sophisticated in vitro systems to recreate 3D tissues and their natural perfusion, the use of animal models is still crucial to preclinically test the potential of immune-based anti-cancer therapies. Considering heterogeneity and complexity of the tumor systems, new models are continuously generated, to reproduce specific types of primary or metastatic malignancies, in the presence of murine or human immune environment. These models are adapted to answer peculiar questions regarding: the effectiveness of specific human immune cell subsets in controlling primary tumor growth or metastasis diffusion, the immune infiltration and the effects of tumor microenvironment, and, also, the relevance of key molecules involved in tumor cell recognition or immune suppression. In this chapter we present a model of human Non-small cell lung cancer (NSCLC) in NSG mice, which lack mature B, T, and NK cells, and are optimal recipient for the infusion and the study of human immune cells, including NK cells. In particular we describe analytical approaches of digital pathology and flow cytometry to characterize and quantify the dissemination of the tumor formations, and the activity of exogenous NK cells in terms of infiltration of the tumor tissue and control of tumor growth and spread.