title: Purification and immunophenotypic characterization of murine MZ and T2-MZP cells.
abstract: B cells are generated every day in the bone marrow, but only a small fraction integrates the peripheral B-cell pool. In the murine spleen, we can find several B-cell subsets representing various maturation stages and/or cell functions. The spleen is a complex lymphoid organ organized in two main structures with different functions: the red and white pulp. The red pulp is flowed with blood while the white pulp is organized in primary follicles, with a B-cell area composed of follicular B cells and a T-cell area surrounding a periarterial lymphatic sheath. The frontier between the red and white pulp is defined as the marginal zone and contains the marginal zone B cells. Because B cells, localized in different areas, are characterized by distinct expression levels of B-cell receptor (BCR) and other surface markers, splenic B-cell subsets can be easily identified and purified by flow cytometry analyses and cell sorting (FACS).Here, we will focus on marginal zone B cells and their precursors giving some experimental hints to identify, generate, and isolate these cells. We will combine the use of FACS analysis and confocal microscopy to visualize marginal zone B cells in cell suspension and tissue sections, respectively.
authors.affiliation: Immunology Unit, Research Center Ospedale Pediatrico Bambino Gesù, IRCSS, Piazza S. Onofrio 4, 00165, Rome, Italy, manuelaanjosm.rosado@gmail.com.
authors.forename: M Manuela
authors.initials: MM
authors.lastname: Rosado
authors.forename: Marco
authors.initials: M
authors.lastname: Scarsella
authors.forename: Simona
authors.initials: S
authors.lastname: Cascioli
authors.forename: Ezio
authors.initials: E
authors.lastname: Giorda
authors.forename: Rita
authors.initials: R
authors.lastname: Carsetti
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D016175:B-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology*; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D016130:Immunophenotyping / Q000379:methods*; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D011947:Receptors, Antigen, B-Cell / Q000032:analysis; D013154:Spleen / Q000166:cytology*
chemical_list: D015703:Antigens, CD; D011947:Receptors, Antigen, B-Cell