Pasut Alessandra, Oleynik Paul, Rudnicki Michael A
Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada.
Methods Mol Biol. 2012;798:53-64. doi: 10.1007/978-1-61779-343-1_3.
Satellite cells are a heterogeneous population of muscle progenitors with stem cell properties responsible for the regeneration of adult skeletal muscle. Increasing interest in the therapeutic potential of satellite cells has challenged researchers with the need to purify a homogenous population of muscle progenitors. Here we provide a detailed protocol for the isolation of a pure population of satellite cells using fluorescence activated cell sorting. We give specific guidelines to ameliorate the reproducibility of the satellite cell isolation protocol with the goal to standardize procedures across labs. This protocol identifies satellite cells within adult skeletal muscle as an enriched population of Integrin α7(+)/CD34(+) double positive cells and CD45, CD31, CD11b, and Sca1 negative (Lin(-)) cells (Integrin α7(+)/CD34(+)/Lin(-)). Functional assay shows that Integrin α7(+)/CD34(+)/Lin(-) satellite cells possess high myogenic potential and ability to regenerate muscle depleted satellite cells upon transplantation.
卫星细胞是具有干细胞特性的异质性肌肉祖细胞群体，负责成年骨骼肌的再生。对卫星细胞治疗潜力的兴趣日益增加，这就要求研究人员纯化出同质的肌肉祖细胞群体。在此，我们提供了一份使用荧光激活细胞分选技术分离纯卫星细胞群体的详细方案。我们给出了具体指南，以提高卫星细胞分离方案的可重复性，目标是使各实验室的操作标准化。该方案将成年骨骼肌中的卫星细胞鉴定为整合素α7(+)/CD34(+)双阳性细胞和CD45、CD31、CD11b及Sca1阴性（Lin(-)）细胞（整合素α7(+)/CD34(+)/Lin(-)）的富集群体。功能分析表明，整合素α7(+)/CD34(+)/Lin(-)卫星细胞具有高成肌潜力，并且在移植后具有再生肌肉中耗尽的卫星细胞的能力。
