Purification and cell-surface marker characterization of quiescent satellite cells from murine skeletal muscle by a novel monoclonal antibody.
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.
Fukada
So-ichiro
S
Department of Immunology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan.
Higuchi
Saito
S
Segawa
Masashi
M
Koda
Ken-ichi
K
Yamamoto
Yukiko
Y
Tsujikawa
Kazutake
K
Kohama
Yasuhiro
Y
Uezumi
Akiyoshi
A
Imamura
Michihiro
M
Miyagoe-Suzuki
Yuko
Y
Takeda
Shin'ichi
S
Yamamoto
Hiroshi
H
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
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D015415:Biomarkers; D015820:Cadherins; C070896:M-cadherin