title: Characterization by mass cytometry of different methods for the preparation of muscle mononuclear cells.
abstract: Biological processes that are mediated by cell-cell interactions in heterogeneous populations are best approached by methods that have single cell resolution. Most of these methods rely on the preparation, from solid tissues, of cell suspensions by enzymatic digestion, followed by analysis of single cell reactivity to an antibody panel that allows the discrimination of cell populations and characterization of their activation state. Thus for any specific biological problem, both efficient and at the same time mild, protocols for cell separation, together with tissue specific panels of antibodies, need to be developed and optimized. Here we characterize an antibody panel that permits the discrimination of mononuclear muscle cell populations by mass cytometry and use it to characterize the cell populations obtained by three different cell extraction procedures from muscle fibers. We show that our panel of antibodies, albeit limited and incomplete, is sufficient to discriminate most of the mononuclear muscle cell populations and that each cell extraction method yields heterogeneous cell populations with a different relative abundance of the distinct cell types.
authors.affiliation: Department of Biology, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Filomena
authors.initials: F
authors.lastname: Spada
authors.affiliation: Department of Biology, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Fuoco
authors.affiliation: Department of Biology, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Pirrò
authors.affiliation: Department of Biology, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Paoluzi
authors.affiliation: Department of Biology, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Castagnoli
authors.affiliation: Department of Biology, University of Rome Tor Vergata, Rome, Italy. Electronic address: cesare.gargioli@uniroma2.it.
authors.forename: Cesare
authors.initials: C
authors.lastname: Gargioli
authors.affiliation: Department of Biology, University of Rome Tor Vergata, Rome, Italy. Electronic address: cesareni@uniroma2.it.
authors.forename: Gianni
authors.initials: G
authors.lastname: Cesareni
mesh_terms: D000818:Animals; D000906:Antibodies; D001709:Biotechnology; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000276:immunology; D018482:Muscle, Skeletal / Q000166:cytology* / Q000276:immunology; D032448:Myoblasts, Skeletal / Q000166:cytology / Q000276:immunology; D059010:Single-Cell Analysis / Q000379:methods
chemical_list: D000906:Antibodies