title: Purification and cell-surface marker characterization of quiescent satellite cells from murine skeletal muscle by a novel monoclonal antibody.
abstract: A novel monoclonal antibody, SM/C-2.6, specific for mouse muscle satellite cells was established. SM/C-2.6 detects mononucleated cells beneath the basal lamina of skeletal muscle, and the cells co-express M-cadherin. Single fiber analyses revealed that M-cadherin+ mononucleated cells attaching to muscle fibers are stained with SM/C-2.6. SM/C-2.6+ cells, which were freshly purified by FACS from mouse skeletal muscle, became MyoD+ in vitro in proliferating medium, and the cells differentiated into desmin+ and nuclear-MyoD+ myofibers in vitro when placed under differentiation conditions. When the sorted cells were injected into mdx mouse muscles, donor cells differentiated into muscle fibers. Flow cytometric analyses of SM/C-2.6+ cells showed that the quiescent satellite cells were c-kit-, Sca-1-, CD34+, and CD45-. More, SM/C-2.6+ cells were barely included in the side population but in the main population of cells in Hoechst dye efflux assay. These results suggest that SM/C-2.6 identifies and enriches quiescent satellite cells from adult mouse muscle, and that the antibody will be useful as a powerful tool for the characterization of cellular and molecular mechanisms of satellite cell activation and proliferation.
authors.affiliation: Department of Immunology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: So-ichiro
authors.initials: S
authors.lastname: Fukada
authors.forename: Saito
authors.initials: S
authors.lastname: Higuchi
authors.forename: Masashi
authors.initials: M
authors.lastname: Segawa
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Koda
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Kazutake
authors.initials: K
authors.lastname: Tsujikawa
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kohama
authors.forename: Akiyoshi
authors.initials: A
authors.lastname: Uezumi
authors.forename: Michihiro
authors.initials: M
authors.lastname: Imamura
authors.forename: Yuko
authors.initials: Y
authors.lastname: Miyagoe-Suzuki
authors.forename: Shin'ichi
authors.initials: S
authors.lastname: Takeda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000954:Antigens, Surface / Q000032:analysis*; D015415:Biomarkers / Q000032:analysis; D015820:Cadherins / Q000032:analysis; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry; D007150:Immunohistochemistry; D051379:Mice; D008815:Mice, Inbred Strains; D018485:Muscle Fibers, Skeletal / Q000166:cytology; D018482:Muscle, Skeletal / Q000166:cytology*; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000276:immunology* / Q000637:transplantation
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D015415:Biomarkers; D015820:Cadherins; C070896:M-cadherin