Purification and cell-surface marker characterization of quiescent satellite cells from murine skeletal muscle by a novel monoclonal antibody
Purification and cell-surface marker characterization of quiescent satellite cells from murine skeletal muscle by a novel monoclonal antibody
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DOI：
10.1016/j.yexcr.2004.02.018
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发表时间：
2004-06-10
影响因子：
3.7
通讯作者：
Yamamoto, H
中科院分区：
文献类型：
--
作者：
Fukada, S;Higuchi, S;Yamamoto, H
本文报道了一种新的抗小鼠肌卫星细胞单克隆抗体SM/C-2.6。SM/C-2.6检测骨骼肌基膜下的单核细胞，并且细胞共表达M-钙粘蛋白。单纤维分析显示，附着在肌纤维上的M-钙黏着蛋白（+）单核细胞被SM/C-2.6染色。从小鼠骨骼肌中新鲜分离纯化的SM/C-2.6（+）细胞在体外增殖培养液中分化为MyoD（+）细胞，在体外分化条件下分化为结蛋白（+）和核-MyoD（+）肌纤维。当将分选的细胞注射到mdx小鼠肌肉中时，供体细胞分化成肌纤维。流式细胞仪分析显示，SM/C-2.6+细胞的静止卫星细胞为c-kit（-）、Sca-1（-）、CD 34（+）和CD 45（-）。此外，SM/C-2.6+细胞几乎不包括在侧群中，但在Hoechst染料外排测定中包括在主要细胞群中。这些结果表明，SM/C-2.6识别和丰富的静止卫星细胞从成年小鼠肌肉，和抗体将是有用的卫星细胞活化和增殖的细胞和分子机制的表征的一个强大的工具。(C)2004年爱思唯尔公司All rights reserved.
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-cadberin(+) 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. (C) 2004 Elsevier Inc. All rights reserved.