title: Purification of Immune Cell Populations from Freshly Isolated Murine Tumors and Organs by Consecutive Magnetic Cell Sorting and Multi-parameter Flow Cytometry-Based Sorting.
abstract: It is well established that tumors evolve together with nonmalignant cells, such as fibroblasts, endothelial cells, and immune cells. These cells constantly entangle and interact with each other creating the tumor microenvironment. Immune cells can exert both tumor-promoting and tumor-protective functions. Detailed phenotypic and functional characterization of intra-tumoral immune cell subsets has become increasingly important in the field of cancer biology and cancer immunology. In this chapter, we describe a method for isolation of viable and pure immune cell subsets from freshly isolated murine solid tumors and organs. First, we describe a protocol for the generation of single-cell suspensions from tumors and organs using mechanical and enzymatic strategies. In addition, we describe how immune cell subsets can be purified by consecutive magnetic cell sorting and multi-parameter flow cytometry-based cell sorting.
authors: Division of Immunology, Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.
authors: Camilla
authors: C
authors: Salvagno
authors: Division of Immunology, Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands. k.d.visser@nki.nl.
authors: Karin E
authors: KE
authors: de Visser
mesh_terms: D000818:Animals; D015415:Biomarkers; D004195:Disease Models, Animal; D005434:Flow Cytometry / Q000379:methods; D018189:Immunomagnetic Separation / Q000379:methods; D016130:Immunophenotyping; D051379:Mice; D009369:Neoplasms / Q000276:immunology* / Q000473:pathology*; D010641:Phenotype; D059016:Tumor Microenvironment / Q000276:immunology
chemical_list: D015415:Biomarkers
keywords: Enrichment; Fluorescence-activated cell sorting (FACS); Immune cells; Magnetic cell sorting; Myeloid cells; Preclinical mouse model; Solid tumor; Tumor microenvironment